Register by March 6th and SAVE! · the westin copley place boston the essential protein engineering summit PEGSFifth Annual final agenda April 6-10, 2009 PPhage Display of Antibodies

Cambridge Healthtech Insti tute250 First Avenue, Suite 300 • Needham, Massachusett s 02494 Telephone: 781-972-5400 Toll-free in the U.S. 888-999-6288 • Fax: 781-972-5425

PEGSummit.com

the westin copley place boston

the essential protein engineering summit

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Phage Display of Antibodies and PeptidesPhage Display of Antibodies and Peptides

Diffi cult to Express ProteinsDiffi cult to Express Proteins

Pre-Clinical / Clinical Development of Pre-Clinical / Clinical Development of Therapeutic AntibodiesTherapeutic Antibodies

Recombinant AntibodiesRecombinant Antibodies

Engineering Protein Therapeutics for DeliveryEngineering Protein Therapeutics for Delivery

Immunogenicity of Therapeutic BiologicsImmunogenicity of Therapeutic Biologics

Monoclonal AntibodiesMonoclonal Antibodies

Protein Scale-Up & ManufacturingProtein Scale-Up & Manufacturing

Aggregation in Protein TherapeuticsAggregation in Protein Therapeutics

Register by March 6th and SAVE!

Premier Sponsor:

KEYNOTE PRESENTERS:

David Blakey, Ph.D., Chief Scientist, Oncology, AstraZeneca/MedImmune

Mark Levick, M.D., Ph.D., Global Head, Biologics Unit, Novartis Institutes for BioMedical Research (NIBR)

Andrew Murphy, Vice President, Target Discovery, Regeneron Pharmaceuticals Inc

Paul W.H.I. Parren, Senior Vice President, Research & Pre-Clinical Development, Genmab

Matthew Robinson, Ph.D., Associate Member, Fox Chase; Researcher, Fox Chase Head and Neck Cancer Keystone Program

George Smith, Ph.D., Curators Professor of Biological Sciences, University of Missouri

John Kulman, Ph.D., Assistant Member, Research Division, Puget Sound Blood Center

Steven J. Swanson, Ph.D., Executive Director, Clinical Immunology, Amgen Inc.

Salvador Ventura, Ph.D., Associate Professor, Institute of Biotechnology, University of Barcelona

George M. Whitesides, Ph.D., Woodford L. & Ann A. Flowers University Professor, Departments of Chemistry & Chemical Biology, Harvard University

Corporate Sponsors:

Corporate Support Sponsor:

TM

HOTEL AND TRAVEL INFORMATIONThe Westin Copley Place10 Huntington AvenueBoston, MA 02116Phone: 617-262-9600Fax: 617-424-7483

Discounted Room Rate: $249 s/dDiscounted Room Rate Cut-off Date: March 13, 2009

Visit PEGSummit.com/travel.asp to book your reservation online or call the hotel directly, (617) 262-9600, to make your room reservation. Please identify yourself as a Cambridge Healthtech Institute conference attendee to receive the reduced room rate. Reservations made after the cut-off date or after the group room block has been fi lled (whichever comes fi rst) will be accepted on a space-and-rate-availability basis. Rooms are limited, so please book early.

Flight Discounts:To receive a 5% or greater discount on all American Airline fl ights please use one of the following methods: Call 1-800-433-1790 (authorization code A4819SS). Go online at www.aa.com (enter A4819SS in promotion discount box). Contact our designated travel agent, Wendy Levine, at 1-800-336-5248 ext. 137. Car Rental Information:Special discount rentals have been established with AVIS for this conference. Call AVIS directly at 800-331-1600 and you must reference our Avis Worldwide Discount (AWD) Number J868190.

Monday 4/6 Tuesday 4/7 Wednesday 4/8 Thursday 4/9 Friday 4/10DISCOVERY

BIOPROCESS

CLINICAL

Phage Display Phage Display Recombinant Recombinant Monoclonal Antibodies Antibodies Antibodies

Diffi cult to Express Diffi cult to Express Proteins for Delivery Proteins for Delivery Protein Scale-Up Proteins Proteins and Manufacturing

Pre-Clinical/Clinical Pre-Clinical/Clinical Immunogenicity of Immunogenicity of Aggregation in Development of Development of Therapeutic Biologics Therapeutic Biologics Protein Therapeutics Therapeutic Antibodies Therapeutic Antibodies

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BIOPROCESS

CLINICAL

Phage Display Phage Display Recombinant Monoclonal Monoclonal Antibodies

Antibodies Antibodies

Diffi cult to Express Diffi cult to Express Proteins for Delivery Protein Scale-Up Protein Scale-Up Proteins Proteins and Manufacturing and Manufacturing

Pre-Clinical/Clinical Pre-Clinical/Clinical Immunogenicity of Aggregation in Aggregation in Development of Development of Therapeutic Biologics Protein Therapeutics Protein Therapeutics Therapeutic Antibodies Therapeutic Antibodies

Register for the suggested PEGS Stream that best relates to your fi eld of work:

DISCOVERY: Phage Display Recombinant Antibodies Monoclonal AntibodiesBIOPROCESS: Diffi cult to Express Proteins for Delivery Protein Scale-Up and ManufacturingCLINICAL: Pre-Clinical/Clinical Development of Therapeutic Antibodies Immunogenicity of Therapeutic Biologics Aggregation in Protein Therapeutics


A p r i l 6 - 1 0 , 2 0 0 9 EVENT-AT-A-GLANCE

Dinner, Presentation and Interactive Panel DiscussionFocus on Aggregation: Mechanisms, Analytical Methods, and Real-World Examples

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SPONSORSHIP OPTIONSSponsored Presentations: Includes a 15 or 30 minute presentation as part of the main conference program

Breakfast or Luncheon Presentation: 30-minute presentation to all attendees in the session room

Both of these opportunities are program-specifi c, allowing you to reach your target audience.

Invitation-Only VIP Functions: Select delegates from the pre-registration list for a private dinner or hospitality suite. CHI will invite the attendees, execute the logistics for the dinner, and conduct all follow-up.

ADDITIONAL OPPORTUNITIES• Conference Tote Bags• Tote Bag Inserts• Literature Chair Drops• Travel Coffee Mugs

• Hospitality Suites• Exhibit Hall Reception• And more!

CHI can customize a sponsorship to meet with your needs and budget. We offer comprehensive packages that give your company exposure before, during and after the event. Sponsorships include a podium talk, exhibit space, conference registrations, branding, use of event mailing lists and more.

AjinomotoAragen BioscienceAttanaAvecia Biologics Blue Sky BiotechConstant SystemsCrown BiosciencesCytovance BiologicsDowpharmaEMD BiosciencesForteBioGeneart

GenetixGENEWIZHaematologic Technologies, Inc.Hyaluron Contract ManufacturingIcX NomadicsNew England BiolabsParagon BioservicesPrecision AntibodyProteos, Inc.QIAGENSloning BiotechnologyTTP Labtech

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To fi nd out more about our comprehensive sponsorship and exhibit packages, please contact:Carol Dinerstein – Senior Manager, Business Development • 781-972-5471 • [email protected]

2009 EXHIBITORS (As of November 26, 2008)

SUNDAY, APRIL 5 (SC2) Phage and Yeast Display Libraries and their Screening10:00am-1:00pmThis workshop is meant to bring scientists up to speed on the display technologies covered by the main conference. The workshop will provide an overview of:

Phage display and construction of phage-displayed peptide, scFv and Fab libraries• Yeast display and construction of yeast-displayed scFv and Fab libraries• Screening • technologies that are compatible with phage- vs. yeast-display libraries

Instructors: Jamie Kathleen Scott, M.D., Ph.D., Professor and Canada Research Chair in Molecular Immunity, Department of Molecular Biology & Biochemistry and Faculty of Health Sciences, Simon Fraser UniversityJames D. Marks, M.D., Ph.D., Professor of Anesthesia and Pharmaceutical Chemistry, Chief of Anesthesia, San Francisco General Hospital, Vice Chairman, Dept. of Anesthesia and Periopera-tive Care, University of California, San Francisco and San Francisco General Hospital

(SC1) Protecting IP for Protein Therapeutics and Diagnostics2:00-5:00pm Sponsored by Morgan, Lewis & BockiusNew rulings continue to emerge from the Supreme Court of the United States that directly impact the ways in which groups and individuals protect their Intellectual Property (IP) in the area of protein therapeutics and diagnostics. Topics to be Covered:

Implications of new supreme court rulings• Strategies for protecting • IPBuilding on existing • IP

Instructors: Sponsored byJohn Iwanicki, Senior Partner, Banner & Witcoff Ltd. Robin Silva, Partner, Morgan, Lewis & Bockius LLP

(SC3) Small versus Large Molecule Therapeutics: Contrasting the Distinct Needs & Requirements for Biologics

2:00-5:00pmAs researchers transition out of working with small molecules and turn to developing biologics, they fi nd they are dealing with numerous challenges that bring a host of distinct needs and requirements that are different than the needs for traditional small molecule pharmaceuticals. From technologies, to facilities, to trial design and the regulations that govern their development, biologics demand appropriate processes and protocols, as compared with drug development. The “Small vs. Large” Short Course will provide a general history and review of the inherent differences between small and large molecule therapeutics as well as provide information on the related issues of development and regulation. Instructors:Barbara Mounho, Ph.D., DABT, Scientifi c Director, Toxicology, AmgenAndrea B. Weir, Ph.D., DABT, Senior Scientifi c Advisor, Navigator Services, Charles River LaboratoriesKenneth Hastings, Ph.D., Associate Vice President, Regulatory Policy, Sanofi Aventis Group

(SC4) Translational Strategies for Development of Monoclonal Antibodies from Discovery to the Clinic

2:00-6:00pmThe effective information fl ow and translation of accumulated knowledge across various antibody development stages remain a major challenge. Successful strategies for development of monoclonal antibodies require integration of relevant knowledge with respect to target antigen properties, antibody design criteria such as affi nity, isotype selection, pharmacokinetic (PK)-pharmacodynamic (PD) properties, and antibody cross-reactivity across species from the early stages of antibody development. Biophysical measurements are one of the critical components necessary for the design of effective translational strategies for lead selection and evaluation of the relevant animal species for preclinical safety and effi cacy studies. Incorporation of effective translational strategies from the early stages of the antibody development process is a necessity; when considered it not only reduces development time and cost, but also fosters implementation of rational decision making throughout all phases of antibody development.Instructors:Mohammad Tabrizi, Ph.D., Director, Global PK-PD and Bioanalysis, MedImmune, Hayward CAGadi Bornstein, Ph.D., Principal Scientist, AstraZeneca R&D BostonScott L. Klakamp PhD, Research Fellow, Biophysical Chemistry and ResearchInformatics, Takeda San Francisco

(SC6) Testing for Immunogenicity2:00-5:00pmDifferent types of bioanalytical assays need to be developed and validated to support the development of therapeutic proteins. This short course will discuss issues encountered with the development and validation of bioanalytical assays as well as planning and interpreting immunogenicity testing in preclini-cal and clinical development programs.Instructors: Moderator: David W. Scott, Ph.D., Professor of Surgery, Microbiology/Immunology, Center for Vascular and Infl ammatory Diseases, University of Maryland, School of MedicineMatthew Baker Ph.D., Chief Scientifi c Offi cer, Antitope, Ltd. Harald Kropshofer, Ph.D., Global Coordinator Immunosafety, F. Hoffmann La Roche AG Mauricio Maia, Ph.D., Scientist, Genentech

(SC7) Next Generation Sequencing Technologies for Antibody Clone Screening Sponsored by Roche/454 Sequencing 2:00 – 5:00 pm

Combining Sequencing With Phage Selections• Single Cell Gene Expression Profi ling• Selection of Clones Using Next Generation Sequencing •

Next Generation Sequencing Technologies for Transcriptome Sequencing Clotilde Teiling, Marketing Manager, Roche Applied Science

High Throughput Screening of Phage Display Antibody Libraries Malini Viswanathan, Ph.D., Principal Scientist, Research, Dyax Corp.

Bioinformatics Methods and Computer Programs for Next-Generation Sequencing Data AnalysisGabor Marth, Ph.D., Assistant Professor, Bioinformatics, Boston College

THURSDAY, APRIL 9(SC5) Dinner, Presentation and Interactive Panel DiscussionFocus on Aggregation: Mechanisms, Analytical Methods, and Real-World Examples5:30 – 8:30pmPresenter & Panel Moderator: John Philo, Ph.D., VP, Biophysical Chemistry, Alliance Protein LaboratoriesThe introductory talk will review the types and mechanisms of aggregation, and why characterization methods in addition to SEC are often requested by the regulators. Advanced methods for characterization of aggregation will be described, including analytical ultracentrifugation (AUC) and light scattering techniques (SEC-MALS and DLS). The advantages and disadvantages of these techniques will be discussed, along with some real-world examples and applications. The audience will gain a better understanding of mechanisms of aggregation, how those mechanisms point to prevention, and which analytical methods should be chosen. Thereafter, the audience and panel members will discuss the issues and learn from each others’ challengesInstructors:Yijia Jiang, Ph.D., Principal Scientist, Formulation and Analytical Resources Department, AmgenTudor Arvinte, Ph.D., Professor, University of Geneva; Chairman and Chief Executive Offi cer, Thera-peomic, Inc.Mary Cromwell, Ph.D., Scientist & Senior Group Leader, Early Stage Pharmaceutical Development, Genentech

*Separate Registration Required

SHORT COURSES*

SPONSORSHIP AND EXHIBITOR INFORMATION

Record Attendance in 2008!Don’t miss out on networking with over 800 of the world’s

leading protein engineering scientists and executives!

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Eleventh Annual

Phage Display of Antibodies and PeptidesApproaches for Second Generation Biologics April 6-7

Recommended Short Courses*SUNDAY, APRIL 59:30 – 10:00 am Morning Short Course Registration10:00am – 1:00pm (SC2) Phage and Yeast Display Libraries and their Screening This workshop is meant to bring scientists up to speed on the display technologies covered by the main conference. The workshop will provide an overview of:

Phage display and construction of phage-displayed peptide, scFv and Fab libraries• Yeast display and construction of yeast-displayed scFv and Fab libraries• Screening technologies that are compatible with phage- vs. yeast-display libraries•

Course Instructors: Jamie Kathleen Scott, M.D., Ph.D., Professor and Canada Research Chair in Molecular Immunity, • Department of Molecular Biology & Biochemistry and Faculty of Health Sciences, Simon Fraser UniversityJames D. Marks, M.D., Ph.D., Professor of Anesthesia and Pharmaceutical Chemistry, Chief • of Anesthesia, San Francisco General Hospital, Vice Chairman, Dept. of Anesthesia and Perioperative Care, University of California, San Francisco and San Francisco General Hospital

1:00 – 2:00pm Afternoon Short Course Registration 2:00 – 5:00 pm (SC7) Next Generation Sequencing Technologies for Antibody Clone

Screening Sponsored by Roche/454 Sequencing * Combining Sequencing With Phage Selections * Single Cell Gene Expression Profi ling * Selection of Clones Using Next Generation Sequencing

Next Generation Sequencing Technologies for Transcriptome Sequencing Clotilde Teiling, Marketing Manager, Roche Applied Science Transcriptome sequencing is a fast and affordable strategy to obtain genomic information that direct and regulates protein synthesis in addition to decoding the sequence that is translated into proteins. From protein, to whole transcriptome, to de novo assembly, The GS FLX System enables the possibility of taking your research to a next level thoroughly. Examples of applications to demonstrate the theory of transcriptome sequencing in cancerous samples and splice variant discovery in Human Genome. This new approach opens a path for new studies combining genetic studies with functional genomics to protein coding.

High Throughput Screening of Phage Display Antibody Libraries Malini Viswanathan, Ph.D., Principal Scientist, Research, Dyax Corp.Antibody phage display technology is well established and widely used for rapidly selecting specifi c antibodies against desired targets. Automated and high throughput approaches for the screening and sequencing of phage libraries can be harnessed to identify high effi ciency lead candidates. The key challenges in managing the information from high throughput data sets and potential solutions will be discussed. Specifi c examples of data management from Dyax will be highlighted.

Bioinformatics Methods and Computer Programs for Next-Generation Sequencing Data AnalysisGabor Marth, Ph.D., Assistant Professor, Bioinformatics, Boston CollegeNext-generation technologies have taken over genome re-sequencing, and are being applied for novel transcript discovery, gene expression analysis, transcription binding site identifi cation, as well as epigenetic analysis. The relatively short read length, the vast throughput, and the new applications made it necessary to develop a number of new computer tools for accurate and effi cient analysis of next-generation sequencing data. This presentation will review those technologies that are currently on the market, describes the informatics challenges to process the data, as well as the computer programs that have been written for various data analysis tasks.


MAIN CONFERENCE

4:00 – 6:00 Main Conference Pre-RegistrationMONDAY, APRIL 6

7:00am Registration and Morning Coffee

8:30 Chairperson’s Opening RemarksGregory A. Weiss, Ph.D., Associate Professor, Department of Chemistry, Molecular Biology & Biochemistry, University of California, Irvine

KEYNOTE PRESENTATIONS

8:40 Phage-Antibodies for the Masses George Smith, Ph.D., Curators Professor of Biological Sciences, University of Missouri

9:10 Healthcare and Biopharmaceuticals: A Patient- Focused Partnership

Mark Levick, M.D., Ph.D., Global Head, Biologics Unit, Novartis Institutes for BioMedical Research (NIBR)Advances in the discovery and development of monoclonal antibodies and recombinant proteins has resulted in better therapeutic options for diffi cult-to-treat diseases. Newer approaches in molecular medicine are creating novel opportunities for highly targeted therapies, as well as challenging the more traditional notions of drug development. Successful biological medicines of the future are likely to be those that deliver better outcomes for patients.

9:40 Target Validation via VelociGene, Fully Human Antibody via VelocImmune

Andrew Murphy, Vice President, Target Discovery, Regeneron Pharmaceuticals IncVelociGene technology has revolutionized our ability to manipulate the mouse genome allowing for single base pair resolution combined with mega base capability. We have exploited VelociGene technology to improve upon earlier generation human antibody mice (“HumAb mice”), by precisely exchanging 6 mega bases of mouse immune genes with their human immune gene counter parts. This (VelocImmune) mouse is perhaps the new standard for generating fully human antibodies against targets of interest, and when combined with VelociMab technologies provides perhaps the fastest approach for going from target to fully human antibody in the clinic. The VelocImmune mouse is part of a technology platform that will generate well over half a billion dollars in combined licensing and collaboration revenues.

10:10 Grand Opening Coffee Break in the Exhibit Hall

LIBRARY DESIGN AND ENGINEERING- Expanding on Diversity 11:10 Phylomers: Peptides Derived from Biodiverse Genomes, as

Blockers for Intracellular as Well as Extracellular TargetsRichard Hopkins, Ph.D., Research Manager, Phylogica Phylomers are a new class of peptide derived from genomic fragments of biodiverse archael and bacterial species. These peptides can have high affi nities and high specifi city for targets, even before affi nity maturation or optimisation which can further increase affi nity to the low nanomolar-picomolar range. Phylomer peptides can also exhibit superior functional hit-rates, when compared to randomly derived peptides, possibly due to an evolutionary selection for structure and stability. The majority of Phylomer peptides are not signifi cantly immunogenic, since their small size reduces the likelihood of containing MHC epitopes. Synthesised Phylomers directed against intracellular targets have been shown to function in animal models of ischemia and wound healing of severe burns, when fused to a protein transduction domain. Inhaled Phylomers have also shown in vivo effi cacy. Phylomers can block both extracellular and intracellular targets, which are challenging to block with small molecules or to access with larger proteins such as antibodies or protein-based scaffolds. Watt PM (2006) Nature Biotechnology 24 (2):177-83.

Sponsored By:

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11:40 Use of Phage Display to Develop Therapeutic Antibodies for the Treatment of TTP, a Potentially Fatal Autoimmune Thrombotic Disorder

Don Siegel, M.D., Ph.D., Professor and Vice-Chair, Department of Pathology & Laboratory Medicine, University of Pennsylvania School of MedicineThrombotic thrombocytopenic purpura (TTP) is a potentially fatal clotting disorder caused by autoantibody inhibition of ADAMTS13, a von Willebrand Factor-cleaving metalloprotease. Antibody phage display was used to dissect the pathogenic autoimmune response responsible for this disorder. Over 100 human anti-ADAMTS13 inhibitory scFv’s have been isolated from multiple TTP patients and have led to the development of crossreactive anti-idiotypic reagents and a novel mouse model system in which to test their potential therapeutic effi cacy.

12:10pm Engineering the Filamentous Phage Surface to Alter its Immunogenicity

Jamie Kathleen Scott, M.D., Ph.D., Professor and Canada Research Chair in Molecular Immunity, Department of Molecular Biology & Biochemistry and Faculty of Health Sciences, Simon Fraser UniversityOur lab has altered the immunogenic properties of the fi lamentous phage by altering the biochemical properties of its surface. We removed the outer domains of the pIII protein of fi lamentous phage, and also altered the amino acid sequence of the N-terminal 10 residues of pVIII, thus changing the surface properties of the phage. Antibodies against wild-type phage did not cross react with the altered phage, and vice versa. In addition, for two different peptides, we prepared phage-immunogens that either display a peptide-target as a recombinant fusion to pVIII, or that comprise a synthetic version of the peptide chemically conjugated to the phage surface. These immunogens were compared for their ability to elicit anti-peptide antibodies, and to “focus” the antibody response against the displayed or conjugated peptide. The results of these experiments and use of phage as model immunogens will be discussed.

12:40 Luncheon Presentation I (Opportunity Available)1:10 Luncheon Presentation II (Opportunity Available)1:40 Break

MODELING EFFECTOR FUNCTION2:00 Chairperson’s RemarksK. Dane Wittrup, Ph.D., J.R. Mares Professor, Chemical Engineering & Bioengineering, Massachusetts Institute of Technology

2:05 Modeling Effector FunctionRene Ott, Ph.D., Lab of Molecular Genetics & Immunology, Rockefeller University

2:35 Complement Proteins And PathwaysMichael C. Carroll, Professor, Pediatrics Pathology, Harvard Medical School

3:05 Refreshment Break in the Exhibit Hall

SCREENING ON CELLS AND COMPLEX ORGANISMS 3:45 The Design and Engineering of Fc Heterodimers for the

Production of Bispecifi c Antibodies Wei Yan, Ph.D., Principal Scientist, Protein Science, Amgen Inc.We have modifi ed the CH3 domain of the Fc interface with a few selected mutations so the engineered Fc proteins preferentially form heterodimers. Our engineering approach takes advantage of electrostatic interactions in promoting Fc heterodimer formation and discouraging Fc homodimers and does not directly affect the hydrophobic core of the CH3 domain interface. The successful production of heterodimeric Fc molecules facilitates the construction of bispecifi c antibodies and various heterodimeric Fc fusion proteins.

4:15 Selection of Internalizing Antibodies on Cells for Targeted Drug and Nucleic Acid Delivery

James D. Marks, M.D., Ph.D., Professor of Anesthesia and Pharmaceutical Chemistry, Chief of Anesthesia, San Francisco General Hospital, Vice Chairman, Department of Anesthesia and Perioperative Care, University of California, San FranciscoWe have developed methodologies which allow direct selection of phage antibody libraries on cells to either known or unknown surface receptors. The resulting antibodies bind to cell surface receptors and are internalized, providing a targeting moiety for the intracellular delivery of drugs or nucleic acids.

4:45 Problem Solving Break-Out Sessions5:45 Networking Cocktail Reception in the Exhibit Hall6:45 Close of Day

TUESDAY, APRIL 7

BI-SPECIFIC ANTIBODIES 8:25am Chairperson’s RemarksMichael J. Feldhaus, Ph.D., Senior Director, Antibody Engineering, Adimab Inc.

8:30 Fc-Engineered CD19 x CD32b Antibody for B-Cell Inhibition in Autoimmune Diseases

John Desjarlais, Ph.D., Vice President of Research, Protein Engineering, XencorWe have engineered the Fc domain of an anti-CD19 antibody to dramatically increase its affi nity for the inhibitory Fc gamma receptor CD32b, which plays a critical role in modulating B cell activation in response to autoantigens. The novel bispecifi c agent has potent B cell inhibition activity and can suppress antibody responses in vitro and in vivo.

9:00 Stability Engineered IgG-Like Tetravalent AntibodyBrian R. Miller, Ph.D., Senior Scientist, Protein Engineering, Biogen IdecThis presentation will discuss technology developed by Biogen Idec for engineering stable single-chain Fv domains that serve as building blocks for constructing IgG-like bispecifi c and tetravalent antibodies. By increasing antibody valency towards a B cell antigen expressed on human leukemia cells, a tetravalent antibody was proposed to be a more potent cross-linking agent for inducing tumor cell apoptosis. Stability engineering of the scFv domain enabled high-level production of the tetravalent molecule and in vitro and in vivo studies with this tetravalent antibody will be discussed.

9:30 Using Computational Modeling and Simulation to Guide the Development of Novel Bispecifi c Antibodies Against the ErbB Network

Brian D. Harms, Ph.D., Senior Scientist, Computational Biology, Merrimack Pharmaceuticals, Inc.The ErbB receptor-mediated signaling network is an attractive therapeutic target in oncology. To develop next generation therapeutics against ErbB-driven cancers, we have employed a systems approach coupling quantitative biology with a predictive mechanistic model of ErbB receptor and downstream signaling networks. We will discuss how design insights from this Network Biology approach have guided Merrimack’s preclinical development of novel bispecifi c approaches for targeting this network.

10:00 Coffee Break in the Exhibit Hall10:45 Multi-Specifi c or Multivalent IGG Antibodies with Engineered

Antigen Binding Sites in the Constant Domains of Heavy and /or Light Chains

Florian Rüker, PhD, f-star GmbH, and Professor, Christian Doppler Laboratory for Antibody Engineering, Department of Biotechnology, BOKU – University of Natural Resources and Applied Life Sciences Viennaf-star’s Modular Antibody Technology can be used to engineer additional binding sites into antibodies without changing the natural antibody format. We have developed large surface display libraries of correctly folded immunoglobulin domains randomized in non-CDR-loop positions. Constant domains of both heavy and light chain with engineered antigen binding sites (SMIDs: small modular immunoglobulin domains), may be used both as stand alone therapeutic entities (e.g. Fcab™) and as building blocks for mAb2™, which are IgG molecules designed to have e.g. multi specifi city, improved avidity, improved effector functions and/or pharmacokinetics compared to their parent monoclonal antibodies.

11:15 Selection of Antibodies Against Multispan Membrane Proteins from Phage Display Libraries

Isidro Hotzel, Ph.D., Scientist, Antibody Engineering, Genentech, Inc.Membrane proteins with multiple transmembrane regions have been traditionally diffi cult targets for selection of specifi c binders from phage display libraries. We developed a new method to easily and rapidly obtain enriched mammalian membrane proteins for phage display applications that bypasses the need for protein purifi cation, reducing the time required for membrane protein production and characterization. Antibodies obtained using this method bind native membrane proteins on the surface of mammalian cells.

11:45 ProteOn, a Versatile Platform for Therapeutic Antibody Development Sponsored by

John Corbin, Ph.D., Senior Scientist I, Molecular Interactions & Biophysics, Preclinical Research & Development, XOMA (US) LLCCharacterization of both target protein and antibody mechanism of action (MOA) can facilitate many phases of therapeutic antibody discovery and development, impacting the strategie used for early stage discovery, in vivo proof-of-concept studies, toxicological assessment, clinical trial design and commercial development. Antibody MOA analysis often starts with the development of methods to monitor target activation by protein-protein interactions such as receptor ligand binding and assembly of an activated signaling complex. Surface plasmon resonance (SPR) based instruments such as the ProteOn XPR36 serve as one of the primary tools for elucidating the biophysical interactions that underlie the mechanisms of target and antibody activation. Due to the unique properties of each antibody-target interaction and specifi c design goals, there is a continual need to develop novel assays for each therapeutic antibody project. SPR platforms that offer fl exible assay design and relatively high throughput are essential to meet this demand and address the wide range of molecular interactions involved in the development of therapeutic antibodies. SPR instruments with parallel fl ow cell arrangement allow for comparative side-by-side analysis of multiple antibodies, immobilization conditions, regeneration conditions and antigen concentrations greatly reducing assay development and optimization time. Specifi c examples of SPR based assays used for mechanism of action studies will be presented.

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12:15pm Luncheon Presentation I (Opportunity Available)12:45 Luncheon Presentation II (Opportunity Available)1:15 Break

CASE STUDIES 2:00 Chairperson’s RemarksChairperson: Richard W. Wagner, Ph.D., President and Chief Executive Offi cer, SRU Biosystems, Inc.

2:05 A Potent Anti-Notch1 Antibody Specifi cally Blocks the Ligand Binding Site and Inhibits Multiple Notch Dependent Processes, Including Tumor Growth

Aaron K. Sato, Ph.D., Senior Director, OncoMed Pharmaceuticals, Inc.The Notch family of receptors are recognized to play central roles in the control of cell fate decisions, and have long been suspected of roles in cancer. We report identifi cation of a critical epitope within the Notch receptor family involved in ligand binding and the development of an anti-Notch1 antibody against it. This antibody completely blocks ligand binding to the Notch1 receptor and subsequent signaling. In xenograft studies this antibody demonstrates anti-tumor activity and impacts upon cancer stem cells.

2:35 Targeting GM-CSF-R: Why High Affi nity and Potency MatterMatthew Sleeman, Ph.D., Director of Biology; Respiratory, Infl ammation and Autoimmunity Therapy Area, MedImmuneIn developing new antibody therapies for the treatment of rheumatoid arthritis a detailed understanding of antigen abundance, antibody pharmacokinetics, route of administration as well as target affi nity and neutralisation is required. In this study we describe the isolation of a high affi nity antibody to the GM-CSF receptor alpha chain by a novel combinatorial method of phage display and discuss the signifi cance of these affi nity and potency gains with respect to its in vivo pharmacokinetic and pharmacodynamic activity.

3:05 Introducing the SKi Pro™ System for 3D Biomolecular Interaction AnalysisJohn Ervin, Director, R&D and Engineering, Silicon Kinetics, Inc.Silicon Kinetics, Inc. introduces the SKi Pro™ system for biomolecular interaction analysis. The SKi Pro system utilizes an optical interferometry design and a novel 3D nanoporous silicon biosurface that promises to increase the sensitivity of biomolecular interaction analysis. The SKi Pro can read both fl ow cells (up to 24 individual self-referenced fl ow cells) for real time kinetic characterization, as well as 96 well plates, for label-free affi nity ranking, end point assays, or concentration analyses. The SKi Pro system offers increased sensitivity, throughput, and versatility at a very affordable price. Learn what 3D can do for your application today.

3:20 Sponsored Presentation (Opportunity Available)3:35 Refreshment Break in the Exhibit Hall

ALTERNATIVE DISPLAY SYSTEMS 4:15 mRNA Display Design of High Affi nity Binding Proteins: Biology, Diagnostics, and TherapyRichard W. Roberts, Ph.D., Associate Professor, Chemistry & Chemical Engineering, University of Southern CaliforniaWe are using mRNA display to create highly specifi c, disulfi de-free proteins based on the human fi bronectin protein domain known as 10FnIII. These molecules provide 1) a new route to protein-based therapeutics 2) a general approach to track and visualize endogenous proteins inside cells, tissues, and organisms, and 3) a useful approach for diagnostic and nanotechnology applications.

4:45 Mammalian Surface DisplayDavid Wenyan Shen, Ph.D., Executive Director and Head, Biologics Research and GlycoFi, Merck & Co., Inc.

5:15 End of Phage Display Conference

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Diffi cult to Express ProteinsEffective Solutions for Improving the Expression of “Finicky” Proteins

Fifth Annual

Recommended Short Course*SUNDAY, APRIL 5

1:00pm Afternoon Short Course Registration

2:00 – 5:00 pm (SC1) Protecting IP for Protein Therapeutics and Diagnostics Sponsored by Morgan, Lewis & Bockius

Instructors: Sponsored by

John Iwanicki, Senior Partner, Banner & Witcoff Ltd. Robin Silva, Partner, Morgan, Lewis & Bockius LLPNew rulings continue to emerge from the Supreme Court of the United States that directly impact the ways in which groups and individuals protect their Intellectual Property (IP) in the area of protein therapeutics and diagnostics. Topics to be Covered:

Implications of New Supreme Court Rulings• Strategies for Protecting IP• Building on Existing IP•

This course provides you with the information critical to protecting your biopharmaceutical intellectual property.


4:00 – 6:00 Main Conference Pre-RegistrationMONDAY, APRIL 6

NEW OR IMPROVED EXPRESSION SYSTEMS 7:00am Registration and Morning Coffee8:30 Chairperson’s Opening Remarks

KEYNOTE PRESENTATION

8:40 Getting More from Less: Integration of Micro-scale Mammalian Expression with a Regeneratable Surface Plasmon Resonance Biosensor Platform

John Kulman, Ph.D., Assistant Member, Research Division, Puget Sound Blood Center

The suitability of mammalian cells as hosts for the production of recombinant human proteins is often counterbalanced by relatively low protein yields, attendant diffi culties in purifying the target from complex media, and bottlenecks in clone screening and expansion. We have developed a novel mammalian cell expression system that circumvents these limitations for the production of ligands suitable for surface plasmon resonance (SPR) studies. Following simple transient transfection, target ligands are purifi ed on-chip from complex serum-containing media and remain stably bound throughout analyte binding and dissociation phases. Residual ligand is then quantitatively removed from the regeneratable chip surface prior to the next capture step. This methodology requires microliter amounts of unfractionated culture medium per binding experiment and enables the sequential analysis of multiple ligand-analyte pairs on a single sensor chip.

9:10 Improving Integral Membrane Protein Production in the Baculovirus Expressions System

Suzanne M. Thiem, Ph.D., Department of Biochemistry and Molecular Biology, Michigan State University

We are using model membrane proteins fused with GFP as tools for investigating why integral membrane proteins are diffi cult to express in the baculovirus expression system, and for identifying and developing new cell lines better suited for membrane protein production. A new insect cell line and progress in developing baculovirus vectors designed for enhanced membrane protein production will be presented.9:40 Template-Directed-Assembly of RTK TK Domains Sponsored by

Domains Restores Biologically-Relevant Function Stephen Zolnay, Vice President of Business Development, P.A. Technologies

Receptor tyrosine kinases and other membrane associated kinases are important targets in drug discovery. Use of recombinant kinase domains, from targets which are normally membrane-associated, in early stage HTS efforts often provide unreliable results including false positives, missed leads, and poor selectivity data (IGF-1R, TrkB, Tie2, and ErbB4, as examples). Because these

target proteins normally reside on the cell membrane and utilize the membrane as a scaffold for assembly of complex signaling networks, we have developed a simple, homogenous, in vitro self-assembly approach to create more meaningful assays. Our template-directed self-assembly approach promotes the assembly of functioning protein complexes on a soluble nanometer scale template, so that the recombinant fragments function more like they do inside the cell. Our results indicate that these proteins are more active, more faithfully represent their cellular function, and have altered substrate selectivity when properly assembled, and therefore provide more meaningful results.

9:55 Rapid Recombinant Protein Expression Sponsored byScreening and Production in Insect Cells

Barbara Morris, Senior Scientist, Product Applications, EMD Chemicals, Novagen Brand

The pIEX/Bac™ plasmids are for plasmid or baculovirus- mediated expression in insect cells. An immediate early promoter allows expression directly in Sf9 cells without viral infection, providing an ideal format for screening multiple constructs. For large scale protein production, the pIEx/Bac plasmids are easily recombined into a baculovirus genome.


11:10 Controlling Transgene Expression via Intragenic CpG Dinucleotides

Asli Bauer, Ph.D., Senior Scientist, Molecular Microbiology and Gene Therapy, University of Regensburg

Here we provide evidence that the intragenic content of CpG dinucleotides signifi cantly infl uences the level of transient and stable protein expression in mammalian cells such as 293T or CHO. Whereas a maximal increase of CpG numbers in the open reading frame of several reporter transgenes resulted in the highest expression yields albeit codon quality was reduced, the elimination of CpGs clearly diminished transgene expression both in vitro and in vivo. Recent results suggest an infl uence of intragenic CpG dinucleotides on nucleosome assembly potentially affecting chromatin structure. Deeper insights into CpG-mediated epigenetic mechanisms might serve to develop novel optimization strategies for improved protein production.

11:40 Novel Expression and Production Methods for Diffi cult to Express Proteins in Tetrahymena thermophila

Paul Colussi, Ph.D, Director of Protein Expression, Tetragenetics

Tetrahymena shares many of the attractive production features of other microbial systems and is uniquely suited to the production of diffi cult to express proteins, most notably highly hydrophobic membrane proteins. Key to this feature is a signifi cant metabolic commitment to membrane protein production and a large membrane surface area that is greatly exaggerated by hundreds of cilia. At the core of our expression system is a ribosomal DNA based vector that is targeted to a unique chromosome encoding ribosomal DNA that is formed at 9000 copies per cell. The palindromic nature of the rDNA chromosome allows stable maintenance of up to 18,000 copies of the transgene per cell. Expression is driven from a choice of powerful but tightly regulated metallothionein promoters. In this presentation we will be detailing case studies of recombinant membrane protein production validating the versatility of Tetrahymena as an expression host.

12:10pm A Novel and Cost-Effective Method to Express Antimicrobial Peptides for Therapeutic Use in Escherichia coli

Bettina Bommarius, Ph.D., Research Associate, Pathology, Emory University

The successful recombinant expression of antimicrobial peptides in bacteria with sumo as a protective group is a useful tool to overcome critical shortfalls in the therapeutic use of antimicrobial peptides, since existing methods for peptide synthesis are too expensive to provide the amounts of AMPs needed in clinical use. Potentially, bacterial overproduction with the subsequent 3 step purifi cation using Ni-NTA and RPC could be scaled up for industrial use and make antimicrobials readily available in clinical trials as a alternative to antibiotics in treatment of MDR pathogens.

12:40 Shuffl e™, A Novel E. Coli Strain Tha Sponsored by Can Promote the Formation of Correct Disulfi de Bonds in the Cytoplasm

Mehmet Berkmen, Ph.D., Research Scientist, Gene Expression, New England Biolabs

Disulfi de bond formation is a post-translational modifi cation essential for the

April 6-7

PEGSummit.com7 PEGSummit.com

folding of many proteins. However, Escherichia coli is ill equipped to correctly fold multi-disulfi de bond proteins to high yields. Here we introduce a novel Escherichia coli strain called Shuffl e™ that can correctly oxidize protein within the cytoplasm. Proteins within the cytoplasm of Escherichia coli are maintained in their reduced non-disulfi de bonded state by thioredoxin and glutaredoxin reductases. Proteins which require disulfi de bonds for their folding are therefore restricted to those that are secreted to extracytoplasmic compartments. In gram negative bacteria such as E. coli, a set of Dsb proteins catalyze the formation of disulfi de bonds in the periplasmic compartment. DsbA catalyzes the formation of disulfi de bonds and the chaperone DsbC isomerizes mis-oxidizes substrates into their correctly folded state. Mutant E.coli lacking the two reductases (trxB and gor), with an additional suppressor mutation (ahpC*) which restores viability, allow the formation of stable disulfi de bonds in the cytoplasm. Under these conditions thioredoxins are in their oxidized state, converting them from reductases to oxidases. Proteins which require disulfi de bonds for their folding thus can be oxidized and form stable disulfi de bonds within the cytoplasm. In this presentation we describe a strain which, in addition, expresses the disulfi de bond isomerase DsbC within its cytoplasm. This feature greatly enhances the fi delity of disulfi de bond formation in the cytoplasm, and proteins with multiple disulfi de bonds are correctly oxidized to signifi cantly higher yields. This strain is now commercially available from New England Biolabs as Shuffl e™ and is constructed in E. coli K12 and E. coli B backgrounds. Further developments were engineering for the expression of T7 derived protein expression and toxic proteins. We will present detailed characterization of Shuffl e™ along with yields of protein obtained.

1:10 Luncheon Presentation II (Opportunity Available)

1:40 Break

PROTEIN ENGINEERING APPROACHES2:00 Chairperson’s RemarksDeb Chatterjee, Ph.D., NCI

2:05 Innovative Low Human Homology Affi nity TagsFons Bosman, Ph.D., Formulation Expert, Biologicals, Innogenetics NV

We designed fl exible small (multifunctional) low human homology affi nity tags (LHH-tag) that maintain the benefi t of single-step purifi cation on commercially available resin, and that can resolve these general immunology-related disadvantages. The selection strategy of these LHH-tags, as well as purifi cation- and immunology-related results will be presented and discussed.

2:35 Fusion Proteins in Mammalian ExpressionChristopher Mehlin, Ph.D., Senior Scientist, Protein Science, Amgen

I will present a small study we have done with six diffi cult-to-express proteins. We made a set of vectors which appended three different solubility and expression enhancing tags and screened the resultant fusions via transient transfection in a HT, parallel system. Some of these tags were clear winners, and others didn’t do so well. I will also discuss issues concerning scale-up and cleavage of these tags.


3:45 Comparative Study on Autologous Sponsored byExpression Improvement in Human Cells by Gene Optimization: Results and Applications

Stephan Fath, Ph.D., Scientist, Research & Development, GENEART AG

We report the largest gene expression study on synthetic optimized genes in mammalian cells to date. Fifty human genes from the NCBI Entrez database representing different protein classes such as protein kinases, cytokines, membrane proteins and transcription factors, were optimized for increased mRNA half-life and protein expression in human cells. Expressed protein levels in HEK-293T cells were quantifi ed and compared. The results clearly indicate a signifi cant improvement of expression yield with optimized constructs compared to respective wildtype versions. Therefore, gene synthesis is not only a versatile manner to obtain individualized genes but also allows for autologous expression increase in most cases.

4:15 E. Coli Mutants Selected for Improved Protein ExpressionJames Bowie, Ph.D., Professor, Chemistry and Biochemistry, University of California at Los Angeles

We have developed a simple genetic selection for improved protein expression. The selection is powerful enough to isolate host mutations that increase expression. I will describe our progress in obtaining E. coli strains with improved abilities to produce membrane proteins.

4:45 Problem Solving Break-Out Sessions

Table 9 Expression of G Protein-Coupled Receptors for Structural StudiesModerator: Alexei Yeliseev, Ph.D., National Institutes of Health

• How to choose the expression strategy• Stabilization of the recombinant receptors in detergent micelles• Site-specifi c and uniformed labeling for spectroscopic studies

Table 10 Protein Production in Alternative MicrobialSystems- Harvesting High-Hanging Fruit

Moderator: Paul Colussi, Ph.D., Tetragenics, Inc.

• Protists (ciliated or otherwise) as expression Moderators• Codon Optimization in alternative expression systems• High-throughput screening applications for alternative expression Moderators

Table 11 Solutions for Diffi cult Proteins: What to Try First?Moderator: Christopher Mehlin, Ph.D., Amgen

• Refolding from E. coli?• Expression in alternate Moderator?• Genetic optimization?• Fusion partners?

Table 12 The Dirty Little Secrets of Production Cell Lines-Rearrangements and Their In-Depth Analysis

Moderator: Dayou Liu, Ph.D., Amgen

• How rearrangements happened.• Why rearrangements are important for production cell lines• Methodology for rearrangement analysis• Assessment of impact of rearrangements

Table 13 The Quest to Increase Protein Expression: Navigating a Battlefi eldModerator: Mouna Guerfal, University of Ghent

This discussion will focus on different approaches used to manipulate organisms in order to increase the production of heterologous proteins. • Techniques as co-expression of molecular chaperones, altering expression

parameters such as growth temperature, supplementing culture medium with chemicals, etc.

• Even though these manipulations can lead to increased expression levels, most of them cannot be used as a general technique to improve the yield of heterologous proteins. Different proteins encounter problems at different levels when expressed in heterologous host strains.

• What more can we do to increase the expression levels of proteins? Which organism is best for which protein? Are we capable to develop routine techniques to increase protein yield? Can the battle for effi cient protein expression be won?

Table 14 Will High Through put protein production solve our problems?Moderator: Marco Casteleijn, Ph.D., University of Oulu

• Introduction – yourself and “agenda”• Proper Protein Folding – pilot versus parallel approaches• High Yield versus Low Volume• The Major bottlenecks in Protein Engineering (and Metagenomics)• Future outlook – what to solve next?

5:45 Networking Cocktail Reception in the Exhibit Hall

TUESDAY, APRIL 7

CASE STUDIES8:25 am Chairperson’s Remarks

8:30 Secreted Expression of Self-assembling Proteins in Pichia Pastoris

Catarina Ferreira da Silva, M.Sc., Bioprocess Engineering, Wageningen University and Research Centre

Custom-made self-assembling proteins, resembling proteins like collagen or silk, may have a broader medical and pharmaceutical application if they could be produced in an animal free way. The optimization of Pichia pastoris as a host organism for the production of these proteins will provide a new system for the synthesis of innovative protein materials with well defi ned conformations and properties.

9:00 “Lost in Translation:” A Study of Human Relaxin 2 and Frameshift Events Associated with the Lysyl-tRNA and the Rare Arginine Codon, AGA

John J. Kerrigan, M.S., Department of Biological Reagents and Assay Development, GlaxoSmithKline

Expression of human proteins in E. coli can be problematic if the mRNA contains codons rarely used by the host cell. In the case of human relaxin 2, frameshift events were observed around the following sequence CGA-AAA-AAG-AGA. This sequence is interesting because it contains both a ‘slippery A sequence’ as well as a rare AGA codon. When human relaxin 2 was expressed in E. coli Bl21(DE3), three molecular weight species were exhibited, one species was the predicted while the other two were mistranslations. Co-expressing a plasmid encoding the cognate arginyl-tRNAArgucu in the host eliminated both of the frameshift events.

88 PEGSummit.com

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Upon registering, you will be notifi ed if you are one of the fi rst 50 to qualify •

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9:30 An Assessment of the Expression of Two Human Therapeutic Proteins in Pfēnex Expression Technology™

Speaker to be Announced

10:00 Coffee Break in the Exhibit Hall

10:45 Expression and Characterization of a Human BMP-7 Variant with Improved Biochemical Properties

Bethany Swencki-Underwood, Ph.D., Senior Associate Scientist, Antibody and Drug Discovery, Centocor Inc.

The pharmaceutical development of recombinant BMP-7 for systemic delivery has presented many challenges. Utilizing structural information, we have designed and generated a number of rational BMP-7 mutations that improved both expression levels in mammalian cells and solubility at neutral pH, while limiting the amino-terminal heterogeneity of the mature protein. Introduction of these mutations did not compromise BMP-7 in vitro bioactivity.

11:15 Parallel High Throughput Expression of a Thermostable Phosphorylase

Marco Casteleijn, Biorpecess Engineering Laboratory, University of Oulu

11:45 Enhancing Yields of Hard to Express Proteins using an Improved Drosophila S2 Expression System

Wian de Jongh Ph.D., Research Scientist, Pharmexa

12:15pm Boost the Innovation Capacity by Integrating Sponsored bywith China Biology CRO

Larry Wang, Ph.D., President, Genscript Corporation

Compared to chemistry, the outsourcing of biological research in drug discovery, is a fairly new phenomenon. However, it has displayed enormous potential for fast growth. GenScript, a leading biology CRO, is already playing a signifi cant role in the development of the fi eld. Spearheaded by seasoned scientists from the pharmaceutical industry, GenScript offers comprehensive services in bio-reagent, assay development and screening, lead optimization, and antibody drug development. In particular, the company has delivered thousands of proteins, solving many diffi cult cases involving protein expression, purifi cation, refolding, and formulation.


1:15 Break

EXPRESSING PROTEINS FOR STRUCTURAL STUDIES2:00 Chairperson’s Remarks

2:05 Expression of Diffi cult, But Medically Important, Proteins for Structural Studies

Opher Gileadi, Ph.D., University of Oxford

2:35 High Resolution Crystal Structure of the Catalytic Domain of ADAMTS-5 (Aggrecanase-2); The Refolding and Crystallization of a “Diffi cult Protein”

Karl Mathis, B.S., Principal Scientist, Primary Pharmacology, Pfi zer, Inc.

Limited expression levels and low solubility of catalytic domain constructs of ADAMTS proteins had previously precluded the determination of a crystal structure of any member of this family. To overcome these obstacles, we used site-directed mutagenesis, high pressure refolding “Barofold PreEMT” technology, and thermal unfolding studies to prepare pure and active catalytic domain of ADAMTS-5 (cataADAMTS-5) from Escherichia coli and determined its crystal structure at 1.4 Å resolution in the presence of an inhibitor. Elucidation of a high resolution three dimensional structure of the catalytic domain of an ADAMTS metalloproteinase enables detailed study of substrate specifi city and the potential for rational design of selective inhibitors of this important class of enzymes.

3:05 Expression of Diffi cult Proteins Using Sponsored byFragment Based Libraries- Application to Protein: Protein Interactions

Eddy Littler, Ph.D., Chief Executive Offi cer, Domainex

The current paradigm in drug discovery demands large amounts of high quality protein for both assays and structural studies. However, in many cases the production of such protein is challenging. This is especially true when the target in demand is a complex protein:protein interaction. In this talk we describe, with examples, the use of Combinatorial Domain Hunting as a powerful method of expressing diffi cult proteins in recombinant systems. We will describe a new technology CDH2 and its use in targets consisting of protein:protein interactions.

3:20 Correlation of Cell-Free and Cellular Protein Sponsored byExpression and the Effect of Gene Optimization: A Large Scale Study

Barbara Maertens, Ph.D., Protein Expression/Proteomics, Qiagen GmbH

Cell-free protein expression in eukaryotic cell lysates allows producing proteins in a fast and economic manner, and proteins can be posttranslational modifi ed. In this study, 50 synthetic wildtype and expression-optimized human genes representing various protein classes (including diffi cult to express membrane proteins) were expressed cell-free and compared to the proteins from cellular expression with respect to expression and functionality. Our results show a signifi cant effect of human gene optimization in cell-free expression and demonstrate through correlation data that cell-free expression is a good screening tool for in vivo protein production.


4:15 Preparation of Functional Human Cannabinoid Receptor CB2 for Structural Studies

Alexei Yeliseev, Ph.D., Staff Scientist, NIAAA, National Institutes of Health

CB2 was expressed in Escherichia coli as a fusion with maltose-binding protein and a decahistidine and StrepTag affi nity tags, and its functional activity confi rmed by ligand binding and G protein activation tests. As much as 2-3 mg of 15N-tryptophan-labeled CB2 could be purifi ed from 1L of culture, and effi cient incorporation of the isotope was confi rmed by mass spectrometry. The receptor was reconstituted into a lipid matrix consisting of monounsaturated phosphatidylcholine and phosphatidylserine Factors required for stabilization of the receptor during detergent solubilization and chromatographic purifi cation were studied. Cholesterol hemisuccinate and cannabinoid agonist CP55940 were identifi ed as the two major factors contributing to stability of CB2 protein in detergent micelles. We further tested a large number of detergents for their ability to (i) effi ciently solubilize lipids; (ii) be removed effi ciently from the proteil-lipid-detergent mixture either by treatment with detergent-absorbing resins or rapid dilution below the critical micellar concentration; (iii) preserve functional activity of the CB2 receptor. The zwitterionic detergent LDAO performed best in solubilization of both lipid and protein and maintaining the structural integrity of the CB2 receptor during reconstitution into liposomes. The ability of CB2-containing proteoliposomes to activate G-proteins in response to agonist binding was studied as a function of detergent, lipid, and CB2 concentration. The lipid-reconstituted, purifi ed CB2 is fully functional as determined by the G protein-coupled assay. The proteoliposomes obtained by either rapid dilution procedure or treatment with ExtractiGel sorbent were shown to be small (120-200 nm), free of residual detergent, and homogenous. These preparations have been shown to perform well in solid-state NMR analysis.

4:45 Addressing the Issues of Protein Expression for Structural Studies: Panel Discussion with Audience Questions

5:15 End of Diffi cult to Express Proteins Conference

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PEGSummit.com 9

Second Annual

Pre-Clinical / Clinical Development of Therapeutic Antibodies

April 6-7Recommended Short Course*

SUNDAY, APRIL 51:00pm Short Course Registration

2:00 – 5:00 pm (SC3) Small versus Large Molecule Therapeutics: Contrasting the Distinct Needs & Requirements for Biologics

As researchers transition out of working with small molecules and turn to developing biologics, they fi nd they are dealing with numerous challenges that bring a host of distinct needs and requirements that are different than the needs for traditional small molecule pharmaceuticals. From technologies, to facilities, to trial design and the regulations that govern their development, biologics demand appropriate processes and protocols, as compared with drug development. The “Small vs. Large” Short Course will provide a general history and review of the inherent differences between small and large molecule therapeutics, as well as provide information on the related issues of development and regulation. Instructors:Barbara Mounho, Ph.D., DABT, Scientifi c Director, Toxicology, Amgen

Andrea B. Weir, Ph.D., DABT, Senior Scientifi c Advisor, Navigator Services, Charles River Laboratories

Kenneth Hastings, Ph.D., Associate Vice President, Regulatory Policy, Sanofi Aventis Group

(SC4) Translational Strategies for Development of Monoclonal Antibodies from Discovery to the Clinic

2:00-6:00pmThe effective information fl ow and translation of accumulated knowledge across various antibody development stages remain a major challenge. Successful strategies for development of monoclonal antibodies require integration of relevant knowledge with respect to target antigen properties, antibody design criteria such as affi nity, isotype selection, pharmacokinetic (PK)-pharmacodynamic (PD) properties, and antibody cross-reactivity across species from the early stages of antibody development. Biophysical measurements are one of the critical components necessary for the design of effective translational strategies for lead selection and evaluation of the relevant animal species for preclinical safety and effi cacy studies. Incorporation of effective translational strategies from the early stages of the antibody development process is a necessity; when considered it not only reduces development time and cost, but also fosters implementation of rational decision making throughout all phases of antibody development.Instructors:Mohammad Tabrizi, Ph.D., Director, Global PK-PD and Bioanalysis, MedImmune, Hayward CAGadi Bornstein, Ph.D., Principal Scientist, AstraZeneca R&D BostonScott L. Klakamp PhD, Research Fellow, Biophysical Chemistry and ResearchInformatics, Takeda San Francisco

*Separate Registration is Required

MAIN CONFERENCE

4:00 – 6:00pm Main Conference Pre-RegistrationMONDAY, APRIL 6

7:00am Registration and Morning Coffee

ADDRESSING THE NEEDS8:30 Chairperson’s Opening RemarksRafael Ponce, Ph.D., Scientifi c Director, Toxicology, Amgen Inc.


8:40 Development of a Portfolio of Antibody Based Therapeutics for Oncology

David Blakey, Ph.D., Chief Scientist, Oncology, AstraZeneca /MedImmune

Medimune have developed a signifi cant preclinical and clinical portfolio of human antibody based oncology projects using both transgenic mouse and display technologies. In this presentation, a selection of the late-stage preclinical/early clinical projects will be presented including MEDI 573 (IGF1/2 ligand) and MEDI 575 (PGFRalpha) projects. The challenges of preclinical evaluation of antibodies which lack mouse cross reactivity will be discussed and examples illustrating the utility of KO/KI transgenic mice and primate PK/PD models will be presented.

9:10 AMG 479, A Fully Human IGF-1R Antibody Therapeutic for Cancer

Frank Calzone, Ph.D., Executive Scientifi c Director, Oncology Research, Amgen, Inc.

9:40 Integrating Modern Biology and Liquid Handling into Conventional Hybridoma Discovery

Fred Kull, Ph.D., Manager, Hybridoma Group, Biologic Reagents and Assay Development, GlaxoSmithKline

Modern hybridoma discovery work integrates recombinant target expression, robotic liquid handling, and exquisite hybridoma selection methods that reveal the richness of the murine antibody repertoire. Various interventions such as choice of strain, target conjugate, use of Bacmam, and robotic selection/characterizations will be illustrated that concertedly improve the effi ciency and robustness of hybridoma work. Examples from the druggable target classes and biomarkers will be given.


ANTIBODY STRATEGIES—Regulations & Planning11:10 The Importance of Planning for Preclinical and Clinical

DevelopmentSuzanne M. Sensabaugh, M.S., M.B.A., Vice President, Regulatory Affairs, Panacea Pharmaceuticals

Prior to devoting resources to preclinical and clinical development of an antibody, a development plan should be put in place. This can be a challenging task, especially for an emerging company with limited resources. This plan should focus on fi nal product labeling and the necessary studies to achieve this labeling. Scientifi c and regulatory considerations and best practices will be discussed.

11:40 Novel Pre-Clinical and Clinical Strategies for Two First-in-Class Antibodies

Mark R. Alfenito, Ph.D., Executive Vice President, Corporate Development, KaloBios Pharmaceuticals, Inc.

To validate our Humaneering™ Technology, which is embedded in both of its clinical leads, and to validate the effi cacy of both of these fi rst-in-class targets, KaloBios has employed novel pre-clinical and clinical strategies. KB001 is an anti-Pseudomonas antibody directed against the PcrV protein of the Type Three Secretion System of Pseudomonas. KB002 and KB003 are chimeric and Humaneered™ antibodies, respectively, directed against Granulocyte Macrophage Colony Stimulating Factor (GM-CSF). Preclinical and clinical strategies and data from both programs will be presented.

12:10pm Species Selection: The Foundation of the Nonclinical Toxicology Program

Marque Todd, Ph.D., Regulatory Strategy Lead, Drug Safety R&D, Pfi zer Inc.

Species selection is a key element in the nonclinical testing strategy for therapeutic antibodies. This presentation will cover the regulatory defi nition of a relevant species and the scientifi c data that must be gathered to support the selection of a relevant species. Situations for which acceptable relevant species are not available will also be addressed.

12:40 Accelerating Therapeutic Protein Development with an Automated, Miniaturized Immunoassay System

Bob Dunst, Ph.D., Field Application Scientist, GYROS


1:40 Break

ANIMAL MODELS--TOXICOLOGY AND SAFETY PHARMACOLOGY 2:00 Chairperson’s RemarksSuzanne M. Sensabaugh, M.S., M.B.A., Vice President, Regulatory Affairs, Panacea Pharmaceuticals

2:05 Preclinical Toxicology Studies to Support the Development of Monoclonal Antibodies: Scientifi c Issues and Challenges

Barbara Mounho, Ph.D., DABT, Scientifi c Director, Toxicology, Amgen

The complex nature of therapeutic monoclonal antibodies gives rise to their

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distinctive characteristics, making these molecules fundamentally different from tradition (small molecule) pharmaceuticals. Thus, in conducting preclinical safety studies for therapeutic monoclonal antibodies, many scientifi c challenges can develop due to the unique properties of these molecules. Additionally, conventional preclinical toxicity testing applied to small molecule pharmaceuticals is often not appropriate for monoclonal antibodies. This presentation will review the types of preclinical toxicology studies that are applicable to monoclonal antibodies, as well as provide an overview of the scientifi c challenges, such as species specifi city and immunogenicity, that can arise when conducting toxicology studies with monoclonal antibodies.

2:35 The Surrogate Approaches in the Development of Monoclonal Antibodies

Mohammad A. Tabrizi, Ph.D., Vice President, Preclinical Development, AnaptysBio

Therapeutic monoclonal antibodies exhibit exclusive specifi city for the target antigen. This unique characteristic discriminates antibodies from other therapeutic modalities such as the traditional small molecule drugs. However, when cross-reactivity of the lead antibody across the species is limited, the antibody development program will require generation of surrogate antibodies or surrogate animal models that are necessary for the conduct of preclinical pharmacology and safety studies. Although the surrogate approach allows examination of pharmaco/toxicodynamic properties in a potentially relevant species, it undoubtedly would enhance the complexities and challenges encountered during the course of antibody development process. During this presentation, the application of surrogate molecules, and surrogate animal models in the development of antibodies will be evaluated.


3:45 Challenges and Opportunities in the Design of Nonclinical Safety Programs to Support First in Human Dosing

Laura Andrews, Ph.D., Vice President, Pharmacology & Toxicology, Genzyme Corp.

Nonclinical development programs that are designed to support the safe clinical use of biotherapeutics, and in particular, those considered to be high risk, have considerations that can be very different from traditional small molecule development programs. These compounds require strategies that use case-by-case scientifi c assessments to create a highly integrated program and rationale that effectively utilizes all available pharmacological, chemical, toxicological, drug disposition and pharmacokinetic knowledge. The objective of this session is to highlight the principles, strategies and applications that are necessary for the successful development of these challenging compounds. Case histories illustrating some unsuccessful and successful examples will be presented. Emphasis will also be placed on how nonclinical studies and techniques can be appropriately used to best determine the risk and ensure the safety for the subjects in the clinical trials that receive new high-risk biotherapeutics.

4:15 First-in-Man Dosing in the Post-TGN1412 Era: Points to Consider in the Development of TRX518, an Anti-human GITR Monoclonal Antibody

Lou Vaickus, Ph.D., Chief Medical Offi cer, ToleRx Inc.

Development programs leading up to a decision strategy on fi rst-in-man (FIM) dosing for monoclonal antibodies (Mab) have become more complicated and extensive in the post-TeGenero world. Some regulatory agencies seem to treat all Mab in the superagonist category and are demanding more justifi cation for FIM doses. However, in many ways the path is easier because it is clearer and we know more now about what to do and not to do. This presentation will be based on experience gained with two Mabs now in the clinic, one in Phase 3 (otelixizumab), and will focus on the considerations for a successful fi rst-in-man dose for TRX518, a novel, humanized, aglycosyl anti-GITR (glucocorticoid-induced tumor necrosis factor receptor family related protein) Mab.

4:45 Problem Solving Break-Out Sessions

Penetration of Biologics into Tumors - Does Size Matter?Moderator: David Blakey, Ph.D., Chief Scientist, Oncology, AstraZeneca Pharmaceuticals

Discuss impact of size and exposure time of biologics on tumor penetration• Discuss impact of target distribution and properties of target such as • internalisation on distribution in tumorsDiscuss impact of affi nity of biologic for target and tumor penetration•

The Utility of Neutralizing Antibody Data in Interpreting Nonclinical Study ResultsModerator: Rafael A Ponce, Ph.D., Scientifi c Director, Toxicology, Amgen Inc.

In vivo• tox/PD dataResults of an • ex vivo drug activity assayData from an• ex vivo neutralizing antibody assay

Protein Manufacturing in Disposable SystemsModerator: Robin Ng, Ph.D., Senior Bioengineer, Cell Culture Process Development, Shire Human Genetic Therapies

Which systems are available?•

Scale-up in disposable systems?• Issues associate• d with protein manufacturing in disposable systems

5:45 Networking Cocktail Reception in the Exhibit Hall

6:45 Close of DayTUESDAY, APRIL 7

PHARMACOKINETICS & PHARMACODYNAMICS8:25am Chairperson’s RemarksRon Gulka, Director, Bioinstrumentation Sales and Marketing, ICx Nomadics

8:30 Important Aspects of PKPD and PD-Biomarkers in Guiding Drug Development

Paul J. Fielder, Ph.D., Senior Director, Pharmacokinetic, Pharmacodynamic & Bioanalytical Sciences, Genentech, Inc. The pharmaceutical development of biological therapies is a complex process, which involves the interplay between the pharmacology and biology of both the target and the biologic. These inherent complexities impact molecule selection, preclinical testing strategies and development of initial clinical plans. This talk will cover the use of PK, PD-biomarkers and PKPD modeling in helping to guide the development of biologics. A major focus will be on how to use these novel approaches to inform key decision points during the development process.

9:00 Evaluation of PK/PD Models for Antibodies Exhibiting Target Mediated Disposition

Ellen Q. Wang, Ph.D., Senior Principal Scientist, Pharmacokinetics & Dynamics & Metabolism, Pfi zer Inc.

The behavior of four PK/PD models for antibodies exhibiting target mediated disposition was characterized using panitumumab as the model drug . Our results indicate that the four commonly-used non-linear disposition models were able to describe panitumumab pharmacokinetic data, but these analyses demonstrate that the models differ substantially with regard to relationships between model parameters and drug-receptor binding, predictions of dose-receptor occupation, predictions of the minimally active biological effect level, and predictions of results of multiple dosing.

9:30 Use of Quantitative Pharmacology in the Development of Monoclonal Antibodies

Wendy S. Putnam, Ph.D., Scientist, Genentech, Inc.

Quantitative pharmacology is a multi-disciplinary approach that integrates data about the biological system, drug characteristics, and disease to translate scientifi c discoveries into successful therapeutics. PKPD modeling and simulation is a powerful tool to perform quantitative pharmacology and inform data-driven decisions. This talk will present a case study where quantitative pharmacology was used in the development of a monoclonal antibody to support dose selection and accelerate timelines.


IN SUPPORT OF ANTIBODY DEVELOPMENT 10:45 Design and Interpretation of Nonclinical Safety Studies for

Biotherapeutics in the Presence of Anti-Drug Antibodies: Findings and Recommendations from BioSafe

Rafael Ponce, Ph.D., DABT, Scientifi c Director, Toxicology, Amgen Washington

11:15 Quantitative Analysis of Therapeutic Antibodies in Biofl uids by Mass Spectrometry

Eric Ezan, Ph.D., Head of Laboratory, Institute of Biology Techniques-Saclay, CEA

Because bioanalysis supports preclinical safety studies, there is considerable need for improving bioanalytical methods. Analysis of therapeutic antibodies in biological fl uids is necessary for preclinical studies of pharmacokinetics and toxicity. Mass spectrometry is emerging as an alternative to immunoassay since it offers speed of development, suffi cient sensitivity, and higher specifi city. This talk will cover recent advances and the advantages of mass spectrometry for the quantifi cation of therapeutic antibodies in biofl uids. Immunoanalysis is the current method for the assessment of therapeutic antibody levels in biological fl uids. Although sensitive, this method has disadvantages, such as assay interference by circulating binding proteins or targets and induced antibodies. Its accuracy may therefore be questioned which may be problematic, especially in preclinical studies when bioanalysis is subject to regulatory constraints and the demands of good laboratory practice. Recent analytical developments in the use of mass spectrometry for recombinant proteins and antibodies suggest that this method may be an easy-to-develop alternative of greater specifi city and suffi cient sensitivity.

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11:45 Structure Assessment of Therapeutic Antibodies by Mass Spectrometry: From Screening to Clinical Batches

Alain Beck, Ph.D., Head of Department, Physico-Chemistry, Institut de Recherche Pierre Fabre

This presentation will focus on case studies of antibody structure-function relationship analyses based on state-of-the art Mass Spectrometry methods. Multiple and complementary Mass Spectrometry methods are used at all stages of mAbs discovery, pre-clinical and clinical development: selection of the best antibody-producing clone with the right glycoprofi le; full structural characterization of research leads and clinical candidates; identifi cation of “hot spots” which may be deleterious for stability, pharmacokinetics and pharmacology; comparability assays for formulation, scaling-up and process transfer. Among the latest developments in Mass Spec, case studies will be presented including fi ne characterization of glycans, of mAb microvariants and of antigen-antibody complexes. Altogether these novel structure-function investigation techniques used early in research provide optimized mAb candidates for pharmaceutical and clinical development.

12:15pm Power & Simplicity in Molecular Interaction Studies Teodor Aastrup, Ph.D., CEO, Attana Sponsored by

The Power of Attana QCM – A Tool for Molecular Interactions AnalysisAlexander Kovacs, Ph.D., Global Product Manager, AttanaAttana, a world leading company in life science applications using the Quartz Crystal Microbalance (QCM) technology is now expanding its portfolio with a new generation instrument; extending its capabilities in crude sample analysis and through-put capacity.The instrument presents a versatile platform for molecular interactions analysis producing robust data in both pure and crude samples. Apart from applications such as off-rate screening, detailed kinetics and concentration determination; the excellent performance of the Attana QCM systems also enables challenging experiments such as thermodynamic studies to be accessible at an affordable price. The use of the Attana QCM technology and its proximity independence also enables the study of macromolecules and larger molecular structures.

Quartz Crystal Microbalance Biosensor in Vascular Biology Lipoprotein and Proteoglycan InteractionsLucia D’Ulivo, Laboratory of Analytical Chemistry, University of Helsinki

The retention and modifi cation of Lipoproteins in the vascular wall triggers the immune system, contributing to atherosclerosis and cardiovascular disease. Previous data have shown the involvement of amino acid sequence of ApoB100 to be possibly involved in the former. Here, we have studied the interactions between the amino acid sequence of ApoB100 with the matrix components using the Attana Quartz Crystal Microbalance technology. This allows understanding of the underlying mechanisms contributing to atherosclerosis and cardiovascular disease.


1:15 Break

CARDIOVASCULAR ANTIBODIES2:00 Chairperson’s RemarksFred Kull, Ph.D., Manager, Hybridoma Group, Biologic Reagents and Assay Development, GlaxoSmithKline

2:05 Preclinical Cardiovascular Safety Testing Strategy for Biologics

Hugo Vargas, Ph.D., Director, Investigative Toxicology, Amgen Inc.

Cardiovascular drug safety evaluation is an integral component during the preclinical and clinical development of new medicines. Like small molecules, biologicals are being developed as important new medicines for the treatment of human disease. From a drug safety perspective, biologicals differ remarkably from small molecules, and require a unique approach to toxicity evaluation, especially in the area of cardiovascular safety assessment. This presentation will provide some perspectives and approaches to consider in the preclinical cardiovascular safety testing of biological therapeutics.

2:35 Antibody Therapy Based on Phosphorylcholine for Treatment in Acute Coronary Syndrome

Ola Camber, Ph.D., Vice President, Pharmaceutical Development, Athera Biotechnologies AB

Atherosclerosis and cardiovascular events related to atherosclerosis are strongly associated to infl ammatory activity and it is generally accepted that the immune system is important in atherogenesis. Treatment principles that specifi cally target the immune reaction in atherosclerosis are not presently available. This presentation describes one approach through raising the levels of antibodies that target phosphorylcholine (anti-PC) in individuals at risk, preventing further development of atherosclerosis. Athera´s strategy is to develop a human monoclonal Ab that binds to the PC epitope on oxLDL, thereby blocking macrophage uptake of oxLDL and formation of atherosclerotic plaque. This concept is based on clinical observations that show that low levels of anti-PC is a risk factor for atherosclerosis and atherothrombosis, and further in vitro and in vivo studies.

3:05 Simplifying Automated Surface Plasmon Sponsored byResonance Analysis of Biomolecular Interactions using Continuous Gradient Injection

Ron Gulka, Director of Marketing, ICx Nomadics

Continuous gradient injection is an SPR technique where kinetic and affi nity data can be determined from a single analyte injection. This greatly reduces analysis time as well as lowers reagent consumption and eliminates the need for performing multiple regeneration cycles. Experimental set-up using SensiQ Pioneer’s innovative liquid handling system and application examples will be discussed.

3:20 Sponsored Presentation (Opportunity Available)


ADVANCING ANTIBODIES 4:15 Potent Payload Technology for Antibody-Drug Conjugates with

DNA-Damaging DuocarmycinsVincent de Groot, Ph.D., CLP, Chief Executive Offi cer, Syntarga B.V.

Syntarga’s Potent Payload Technology comprises the combination of highly potent DNA-damaging duocarmycins with sophisticated linker technologies. Several formats have been developed and have been compared with respect to their pharmacological effects in in vitro and in vivo models. We discuss aspects of the chemical design of Potent Payloads as to drug potency, linker stability and therapeutic window of the resulting Antibody-Drug Conjugates (ADCs) and provide an overview of our linker technologies and duocarmycin drug discovery efforts. Preclinical effi cacy and other biological data of Potent Payload-based ADCs will be presented.

4:45 Translational Research in the Development of Novel Nanobody-Based Therapies

Josi Holz, M.D., Chief Medical Offi cer, Ablynx nv

NanobodiesTM are a novel class of antibody-derived therapeutic proteins. Because of their small size, unique structure and extreme stability, NanobodiesTM combine the advantages of conventional antibody therapeutics with the key features of small-molecule drugs. The talk will illustrate the translation of results from the pre-clinical development of the two lead compounds into the clinical development plan to accelerate the advancement of Nanobodies in the relevant therapeutic areas.

5:15 End of Therapeutic Antibodies Conference

Sponsoring Society:

Media Partners:

12PEGSummit.com

Tenth Annual

Recombinant AntibodiesFrom Concept to Clinic

April 8-9Recommended Short Courses*

SUNDAY, APRIL 51:00pm Short Course Registration2:00 – 5:00 pm (SC1) Protecting IP for Protein Therapeutics and

DiagnosticsInstructor: John Iwanicki, Senior Partner, Banner & Witcoff Ltd.New rulings continue to emerge from the Supreme Court of the United States that directly impact the ways in which groups and individuals protect their Intellectual Property (IP) in the area of protein therapeutics and diagnostics. Topics to be Covered:

Implications of New Supreme Court Rulings• Strategies for Protecting IP• Building on Existing IP•

This course provides you with the information critical to protecting your biopharmaceutical intellectual property.

2:00-6:00pm (SC4) Translational Strategies for Development of Monoclonal Antibodies from Discovery to the Clinic

The effective information fl ow and translation of accumulated knowledge across various antibody development stages remain a major challenge. Successful strategies for development of monoclonal antibodies require integration of relevant knowledge with respect to target antigen properties, antibody design criteria such as affi nity, isotype selection, pharmacokinetic (PK)-pharmacodynamic (PD) properties, and antibody cross-reactivity across species from the early stages of antibody development. Biophysical measurements are one of the critical components necessary for the design of effective translational strategies for lead selection and evaluation of the relevant animal species for preclinical safety and effi cacy studies. Incorporation of effective translational strategies from the early stages of the antibody development process is a necessity; when considered it not only reduces development time and cost, but also fosters implementation of rational decision making throughout all phases of antibody development.Instructors:Mohammad Tabrizi, Ph.D., Director, Global PK-PD and Bioanalysis, MedImmune, Hayward CAGadi Bornstein, Ph.D., Principal Scientist, AstraZeneca R&D BostonScott L. Klakamp PhD, Research Fellow, Biophysical Chemistry and ResearchInformatics, Takeda San Francisco *Separate Registration is Required

WEDNESDAY, APRIL 87:30am Registration and Morning Coffee

ANTIBODY AFFINITIES, POLYSPECIFICITY AND SINGLE MOLECULE STUDIES8:30 Chairperson’s Opening Remarks

8:40 Mechanisms of Action of Ofatumumab: A Novel Therapeutic Monoclonal Antibody against CD20

Paul W.H.I. Parren, Senior Vice President, Research & Pre-Clinical Development, Genmab

CD20 monoclonal antibodies represent one of the fl agships of immunotherapy as they continue to revolutionize the treatment of B cell cancers and infl ammatory diseases. Their mechanisms of action have been investigated in detail and this knowledge has been at the basis of the development of ofatumumab. A key mechanism of CD20 antibodies is the killing of target cells via the recruitment and activation of complement. Ofatumumab is a unique human monoclonal antibody that targets a distinct small loop epitope on the CD20 molecule and displays an exceptional effi cacy in recruiting complement and inducing specifi c cell lysis. Novel insights into the mechanisms of tumor cell killing by ofatumumab and its effi cacy in a pivotal clinical trial in B-CLL will be discussed.


9:10 Constant Region Effects on Antibody Specifi city and Affi nityArturo Casadevall, M.D., Ph.D., Chair, Department of Microbiology & Immunology, Albert Einstein College of Medicine Current dogma views immunoglobulins as bifunctional molecules where the variable and constant region function essentially independently. In recent years our laboratory has obtained evidence that variable region identical antibodies differing in constant region manifest different binding characteristics for univalent ligands indicating the existence of constant region effects on specifi city and affi nity. The results have important implications for our views of antibody function, variable region and isotype restriction, and idiotype responses.

9:40 Variable Lymphocyte Receptors as Natural Non-Ig Antigen-Binding Proteins

Roy A. Mariuzza, Ph.D., Center for Advanced Research in BioTechnology, University of Maryland BioTechnology Institute To date, the search for alternatives to antibodies has focused on synthetic binding scaffolds. However, the only natural antigen receptors that are not Ig-based are the variable lymphocyte receptors (VLRs) of lamprey and hagfi sh. X-ray crystallographic analysis of the complex between an anti-protein VLR, isolated by yeast surface display, and its protein antigen has revealed the structural basis for recognition by these novel leucine-rich repeat-based receptors, and its relationship to recognition by conventional antibodies.

10:10 Coffee Break in the Exhibit Hall11:10 The Human Autoantigenome and Predictive AutoantibodiesAbner Notkins, M.D., NIDCR, NIH

11:40 High Affi nity Lamprey AntibodiesZeev Pancer, Ph.D., Assistant Professor, UMDThe antigen receptors of jawless vertebrates are called variable lymphocyte receptors (VLR), and consist of highly diverse leucine-rich repeats. The potential diversity of the sea lamprey VLR repertoire can exceed 1014 different receptors. Yeast surface displayed VLRs from naïve or immunized animals can be used to select high affi nity and high avidity antigen-specifi c binders. VLRs that bind nanomolar concentrations of protein and carbohydrate ligands were isolated and characterized. VLRs are amenable to in vitro mutagenesis to increase affi nity for the ligand. The evolutionary oldest antibodies will soon take a central place in the biotechnology arena.

12:10pm Luncheon Presentation I Slonomics® – A Unique Technology for the Generation of

Highly Designed Gene Libraries Sponsored byThomas Waldmann, Ph.D., Science and Technology Applications, Sloning BioTechnology Sloning specializes in the synthesis of highly genetically diverse and precise customized gene libraries for protein engineering up to 1011 different variants. Unlike traditional methods that rely on single-stranded oligonucleotides, the patented Slonomics® technology uses a set of double-stranded DNA triplets as universal building blocks. Predefi ned triplets represent all possible sequence combinations required to synthesize any gene – ‘one codon at a time’. For library production, multiple codons can be introduced in parallel during the synthesis of a gene construct at any desired sequence position. The absence of functional bias and the ability to select and precisely control delivery up to 20 specifi c codons per sequence position and at any ratio results in exceptionally high quality libraries containing the complete set of desired mutants. In this talk, new technology advancements regarding the random integration of codons into gene sequences and practical examples for improved screening success with SlonoMax™ mutant libraries will be presented.

12:40 Luncheon Presentation II (Opportunity Available)1:10 Break

CASE STUDIES1:30 pm Chairperson’s RemarksLutz Jermutus, Ph.D., MedImmune

1:35 VB6-845: An Immunotoxin Optimized for Clinical DevelopmentJeannick Cizeau, Ph.D., Director, Research, Viventia Biotech Inc.Immunotoxins are comprised of a cell-targeting domain linked to a bacterial or plant-derived cytotoxic payload and represent a highly potent alternative to conventional anti-cancer agents. However, most cytotoxic payloads are inherently immunogenic and thus have limited use therapeutically. VB6-845 is a recombinant fusion protein consisting of a tumor-targeting Fab specifi c for epithelial cell adhesion molecule (EpCAM) linked to bouganin, a plant-derived ribosome-inactivating protein that prevents protein synthesis leading to cell death. The bouganin moiety was de-immunized through the identifi cation and removal of T cell epitopes. The molecular design, purifi cation strategy, biological characterization, and preclinical experience will be presented.

2:05 Engineering Antibody Species Cross-ReactivityDavid Lowe, Ph.D., Head of Display Technology, RI & A, MedImmuneThe development of antibodies as therapeutics for human diseases involves pre-clinical testing in vivo for pharmacological and toxicological assessment. In order to expedite this process, cross-reactivity to non-human homologues of the target antigen is highly desirable. We report here a case-study exemplifying an antibody optimisation project which was designed to both improve affi nity to the human antigen, and additionally to introduce high affi nity binding to pre-clinical animal species.

2:35 Using the Octet QK as a Rapid Screening Sponsored by Platform

Brian Miller, Ph.D., Senior Scientist, Biogen-Idec

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In this presentation I will demonstrate how we’ve incorporated the ForteBio Octet QK into our protein engineering platform. We routinely screen protein libraries expressed in E. Coli for molecules displaying improved biophysical properties, and the Octet allows us to quickly triage based on affi nity measurments on crude samples. This allows us to winnow out molecules which have reduced binding to their targets. In addition, we use the Octet in epitope mapping experiments where we can express large numbers of variant antigens in either E. Coli or yeast systems and determine which substitutions affect antibody affi nity, again without having to scale-up or purify the antigens. In programs where we have multiple candidate Mab’s or Fab’s, the Octet allows us to rapidly preform cross blocking assays in order to group the antibodies in to similar bins. Finally, the Octet has become an indispensible tool for determinng antibody titer for research scale production.

2:50 Sponsored Presentation (Opportunity Available)3:05 Refreshment Break in the Exhibit Hall3:50 Problem Solving Breakout Sessions

Expression of Therapeutic Antibodies for Pre-Clinical TestingModerator: Gerard Casperson, Ph.D., Pfi zer, Inc.• Expression systems/cell lines• Showing comparability from different cell lines• Dealing with diffi cult to express/purify antibodies4:50 Networking Cocktail Reception in the Exhibit Hall6:00 Close of Day

THURSDAY, APRIL 9

BASIC SCIENCE: UNDERSTANDING, SELECTION, EXPRESSION8:30 Chairperson’s Comments8:35 Understanding Agonism Activity of Anti-Fas AntibodiesMatthieu Chodorge, Ph.D., Research Scientist, Oncology Display Technology, MedImmuneDeveloping agonistic antibodies against death receptors is an interesting route to potentially deliver anti-tumor molecules that induce apoptosis of cancerous cells. The talk will fi rst focus on isolation, biochemical characterization, 3D-structure determination and affi nity maturation of agonistic antibodies against Fas death-receptor. In a second part, these results will be discussed to improve our understanding of the agonism activity of anti death-receptor antibodies.

9:05 Selection of Conformational Antibodies and Their Use in Cell Biology

Franck Perez, Ph.D., Research Director, CNRS UMR 144, Institute CurieThe phage display and recombinant antibody approach, while very powerful, is not yet

considered as a valid alternative to classical antibody production in experimental research like in cell biology. Improving both the selection protocols and the antibody production we show that unique tools, sensitive to conformation modifi cation, can be selected by the antibody phage display approach, and bring unexpected information about cell physiology.

9:35 Therapeutic Antibody Production in Mammalian Cells at Lab Scale: Comparison, Adaptation and Implementation of Strategies for Improvements in Quality, Time and Cost

Timothy Oliphant, Ph.D., Biotherapeutics, Pfi zer Inc.We have explored a variety of options for laboratory-scale (up to severalgrams) production of biotherapeutic antibodies from mammalian cells, including large scale transient transfection with PEI and other lipids, baculovirus transduction and production from stably-transfected pools of cells. We have also evaluated process improvements such as fed-batch strategies to increase productivity. We will discuss implementation and impact of improvements that allow rapid production of gram quantities of antibody therapeutics suitable for evaluation in animal models.

10:05 Coffee Break in the Exhibit Hall11:05 Novel Site-Specifi c PEGylation of Antibody FragmentsJi-won Choi, Ph.D., Department Head, Biology, PolyTherics LimitedWhile antibody fragment technology has brought the benefi t of target specifi city of a whole length antibody and the manufacturing advantages of a small protein, they suffer from a short half-life in blood. We have developed a novel method of site-specifi c PEGylation that retains the biological activity of a domain antibody without affecting its tertiary structure. This technology can be applied to a wide range of antibody fragments.

11:35 Spatially Addressed Combinatorial Protein Libraries for Drug Discovery

Vaughn Smider, M.D., Ph.D., Fabrus LLC Antibody discovery is typically accomplished using a display technology (phage, yeast, ribosome, etc.) whereby the gene encoding the antibody fragment and the fragment itself is physically linked. Large pools of displayed antibody fragments are then selected to identify those with the highest affi nity to a target of interest. These methods contrast with small molecule discovery, where combinatorial chemistry libraries are produced in a spatially addressed format, then screened on targets or even in functional cell-based assays. We have developed a high throughput scalable methodology to produce highly pure proteins in spatially addressed format. Thus, recombinant proteins can be produced and screened similarly to small molecule libraries. Libraries of recombinant antibodies have been produced and screened in unique assays not amenable to traditional biologic discovery technology. Hit rates for binding targets are much higher than expected for a naïve antibody library, suggesting that the type of diversity is more important than simply the total number of sequences present in a collection. This paradigm shift in discovery format for biologic discovery could enable identifi cation of unique molecules against new targets, opening certain target classes to antibody therapeutics.

12:05pm Close of Recombinant Antibodies Conference

Fourth Annual

Engineering Protein Therapeutics for DeliveryInvolving Delivery at the Point of Discovery to Produce Best-in-Class Products April 8-9

WEDNESDAY, APRIL 8

IMPROVEMENTS IN HALF-LIFE AND POTENCY 7:30am Registration and Morning Coffee8:30 Chairperson’s Opening RemarksMary Haak-Frendscho, Ph.D. President and CSO, Takeda Pharmaceuticals, San Francisco


8:40 Harnessing Materials for Biologic Drug Delivery George M. Whitesides, Ph.D., Woodford L. & Ann A. Flowers University Professor, Departments of Chemistry & Chemical Biology, Harvard UniversityBi-(and higher) valency is associated with tight binding of proteins to surfaces. This talk will discuss the biophysics of bivalency in molecular recognition by proteins, and speculate on some possible uses.

9:10 Alternative Delivery of NanobodiesHilde Revets, Ph.D., Senior Director Technology, Ablynx nv NanobodiesÒ are the smallest functional fragment of heavy chain only Llama derived monoclonal antibodies. Protein engineering and formatting transforms these NanobodiesÒ in powerful tools in the diagnostic and treatment of various diseases. The inherent stability of the molecules makes them ideal candidates for exploratory studies on alternative ways of drug

delivery through the skin (SC), orally and pulmonary to name a few. The data presented will review the capabilities of NanobodiesÒ administered beyond the intravenous injection route.

9:40 Site-Specifi c PEGylation to Improve the PK/PD Properties of Biologics

Jason Pinkstaff, Ph.D., Associate Director, Preclinical Science, Ambrx, Inc.Ambrx uses an expanded set of amino acids to address the limitations intrinsic to the 20 natural amino acids. Our approach, termed protein medicinal chemistry™, combines the power of medicinal chemistry with recombinant biosynthesis. Through the application of Ambrx’s proprietary technology, numerous variants of the naturally occurring wild-type protein can be generated, with each variant containing a single Ambrx Amino acid incorporated into the protein backbone. A single polyethylene glycol (PEG) molecule can then be attached site-specifi cally to the incorporated Ambrx amino acid. Molecules that vary only in the site of PEG attachment can exhibit signifi cant differences in pharmacology, receptor binding, effi cacy and biophysical stability.

10:10 Coffee Break in the Exhibit Hall11:10 Systemically Acting Chemotherapeutic Enzyme Engineered

for High Activity, Serum Stability and in Vivo Half-LifeGeorge Georgiou, Ph.D., Department of Chemistry and Biochemistry, University of Texas, AustinThis presentation will describe the development and preclinical evaluation of engineered human therapeutic enzymes for amino acid depletion therapy in cancer. Protein engineering approaches were employed to engineer enzymes with high catalytic activity and low immunogenicity. Additionally, we have devised new approaches for increasing the half life of these proteins and for achieving optimal pharmacodynamic profi les for cancer treatment.

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11:40 Oral Delivery, Strongly Reduced Degradation and Strongly Enhanced Effi cacy Resulting from Non-Native Thioether-Bridged Amino Acids, Enzymatically Introduced in Therapeutic Peptides

Gert Moll, Ph.D., Project leader, Biomade Technology FoundationWe stabilized therapeutic peptides by introducing thioether bridges. The resulting peptides can be delivered orally, pulmonarily and subcutaneously, are proteolytically resistant, have up to 100-fold higher in vivo concentration in blood plasma of rats, have strongly enhanced receptor interaction and strongly enhanced therapeutic potential.

12:10 Luncheon Presentation I (Opportunity Available)12:40 Luncheon Presentation II (Opportunity Available)1:10 Break

LOCAL DELIVERY OF AGONISTS 1:30 Chairperson’s RemarksTudor Arvinte, Ph.D., Chairman, Chief Executive Offi cer, Therapeomic Inc. and Department of Pharmaceutics and Biopharmaceutics, School of Pharmacy Geneva-Lausanne, University of Geneva

1:35 Relevance of Transforming Growth Factor Beta 3 (TGF-ß3) for Local Therapies

Tudor Arvinte, Ph.D., Chairman, Chief Executive Offi cer, Therapeomic Inc. and Department of Pharmaceutics and Biopharmaceutics, School of Pharmacy Geneva-Lausanne, University of GenevaOne of the most promising ways to stimulate tissue repair is the application at the injured site of growth factors. TGF-ß3 is an important mediator of growth, maintenance and repair processes in human cells. TGF-ß3 is abundant in bone matrix and was shown to stimulate bone and connective tissue formation as well as the growth of blood vessels. Beside the right choice of the growth factor, the success of local repair therapies is very much dependent on the formulation of the growth hormone formulation and the delivery system. Optimal formulations of TGF-ß3 were developed based on the understanding of the molecules’ biophysical and biochemical properties and by in vivo optimization using relevant animal models. Data will be presented on the use of TGF-ß3 in local bone repair and in the local stimulation of angiogenesis and vasculogenesis.

2:05 Local Delivery of Drugs Into the Eye Using Electroporation Elodie Touchard, Centre de Recherche des Cordeliers, René Descartes University, Pierre et Marie Curie UniversityWe have developed an innovative local protein delivery strategy for the eye. The ciliary muscle is a smooth muscle responsible for accommodation. This strategy is based on the use of ciliary muscle as the target of non viral gene transfection, resulting in an intraocular bioreactor that is able to secrete any therapeutic protein in the ocular media on the long term. Preclinical studies in rat models of ocular diseases will highlight the therapeutic potential of this new concept which may become a useful alternative to conventional current therapies, especially for the management of the most common ocular diseases leading to blindness in patients.

2:35 Thiotransferrin: A Recombinant Human Sponsored byTransferrin Engineered for Site Specifi c Conjugation and Delivery

Joanna Hay, Ph.D., Senior Research Scientist, NovozymesNovozymes has recently developed and manufactured a non-glycosylated human transferrin, the fi rst microbially expressed cGMP grade recombinant transferrin, CellPrimeTM rTransferrin-AF. This recombinant transferrin is expressed in our proprietary yeast expression system which has a long and successful track record in the production of protein therapeutics and has GRAS status with the FDA. Novozymes’ extensive experience in protein engineering has been used to produce novel transferrin variants, ‘Thiotransferrin’, suitable for the site directed conjugation of bioactive compounds. Thiotransferrin possesses an engineered unpaired cysteine; enabling attachment of a bioactive compound at one or more pre-defi ned sites through reaction with the free thiol group of thiotransferrin to form an active therapeutic bioconjugate. Unlike traditional coupling technology which can result in heterogeneous end products; thiotransferrin is designed to allow site-directed conjugation of a bioactive to transferrin resulting in a chemically well defi ned product, necessary for pharmaceutical manufacture. We will present innovative data demonstrating that thiotransferrin is a novel bioactive carrier suitable for targeted delivery to cells expressing the transferrin receptor.

2:50 Sponsored Presentation II (Opportunity Available)3:05 Refreshment Break in the Exhibit Hall3:50 Bio-compatible Polymeric Materials for the Controlled Delivery

of DrugsW. Mark Saltzman, Ph.D., Goizueta Foundation Professor of Biomedical Engineering & Chemical Engineering; and Chair, Department of Biomedical Engineering, Yale University

4:20 Remyelination and Repair of the Central Nerve System by Blocking LINGO-1 Pathway

Sha Mi, Ph.D., Principal Investigator, Neuro- Discovery Biology, Biogen IdecMultiple sclerosis is an infl ammation induced neurodegeneration disease in which axons are demyelinated, degenerated and functional impaired, eventually leading to catastrophic motor function failure. Current therapies mainly focus on blocking the activation and infi ltration of immune T-cells involved in demyelination. There is no therapy available for remyelination and repair of the damaged axons, which is the goal of my laboratory. We have discovered that LINGO-1 is a potent inhibitor for oligodendrocyte differentiation/myelination, and demonstrated that blocking LINGO-1 function by antagonist anti-LINGO-1 antibodies promote OPC differentiation/myelination in vitro and in vivo. Antibodies to LINGO-1 are being developed as a fi rst in class therapeutics for the treatment of MS.

4:50 Networking Cocktail Reception in the Exhibit Hall6:00 Close of Day

THURSDAY, APRIL 9

CNS DELIVERY OF PROTEINS8:30am Chairperson’s RemarksMary Haak-Frendscho, Ph.D., President and CSO, TSF

8:35 A Revolution in CNS Drug Delivery and AssessmentPeter Hoffmann, Vice President, Technology Development, Genzyme PharmaceuticalsDue to a natural defense of the Blood Brain Barrier (BBB), suffi cient quantities of potentially effective therapeutic agents for the treatment of central nervous system (CNS) based pathologies such as Alzheimer’s, Multiple Sclerosis, Stroke and Brain cancer have been unsuccessfully delivered to the site of their required action. Working closely with academic collaborators, Genzyme Pharmaceuticals and Pharmidex have designed CerenseSM to deliver and assess the transfer of a variety of molecules across the BBB.

9:05 Recombinant AAV Delivery of scFv to Modulate the Progression of Prion Disease

Howard J. Federoff, M.D., Ph.D., Executive Vice President for Health Sciences and Executive Dean, Georgetown University Medical CenterThis presentation will advance a preclinical therapeutic strategy for the Prionoses. Delivery of engineered single chain antibodies from a virus vector will be shown to alter the natural history of a mouse prion disease.

9:35 Large Molecule Brain DeliveryClaude Dagenais, B.Pharm., Ph.D. Senior Scientist, Neuroscience Department, Amgen Inc.An overview of the approaches that have been proposed to deliver protein therapeutics to the brain will be presented. Specifi cally, the types of brain delivery vectors, mechanisms of transport, and basic blood-brain transport concepts and methodologies will be discussed.


TUMOR TARGETING SESSION: Smart Pharmacology to Target Specifi c Tissues and Organs 11:00 Chairperson’s RemarksHo Sung Cho, Ph.D., Vice President, Technology & Process Development, Ambrx Inc.

11:05 Clinical Proof of Concept for Bispecifi c Single-Chain Antibodies of the BiTE Class in the Treatment of Cancer

Patrick A. Baeuerle, Ph.D. Professor for Immunology, Senior Vice President, Chief Scientifi c Offi cer, Micromet, Inc.Blinatumomab (MT103/MEDI-538) is a CD19-/CD3-bispecifi c single-chain antibody construct that can potently engage T cells for redirected lysis of normal B and B tumor cells. An ongoing clinical study has shown partial and complete tumor regression in patients with relapsed non-Hodgkin’s lymphoma. A pipeline of new BiTE antibodies is evolving based on fully human and primate crossreactive single-chain antibodies.

11:35 Proving the Benefi t of Antibody-Maytansinoid Conjugates as Anticancer Therapeutics

Peter U. Park, Ph.D., Senior Director, Discovery Research, ImmunoGen, Inc.Even with the success of anticancer antibodies, such as trastuzumab, rituximab and cetuximab, tens of anticancer antibodies have failed to achieve clinical effectiveness, mostly due to insuffi cient therapeutic potency. This is especially true in solid tumors where even the approved mAbs only show modest therapeutic activity when used as a single agent and display their full clinical benefi t in combination with chemotherapeutic agents. Antibody-maytansinoid conjugate is composed of a tumor-targeting monoclonal antibody and a derivative of the potent anti-mitotic microtubule agent, maytansine, which is covalently linked to the antibody. Antibody-maytansinoid conjugates improve the targeted potency of mAbs that do not have enough activity on their own or further enhance the potency of active mAbs. This presentation discusses the pre-clinical evaluation and promising clinical data emerging for antibody-maytansinoid conjugate compounds now in testing.

12:05 End of Engineering Protein Therapeutics for Delivery Conference

Reasons You Should Present Your Research Poster at PEGS:

Your poster will be exposed to over 900 • delegates Receive $50 off your registration: To receive the • $50 discount you must submit your poster title when registering for the meeting Your poster abstract will be published in our • conference CDYour research will be seen by leaders from top • pharmaceutical, biotech, academic and govern-ment institutes

To secure a poster board and inclusion in the conference CD, your abstract must be submitted, accepted,and registration paid in full by March 6, 2009. You will be prompted to submit your abstract electronically when you register for the meeting.

Reasons You Should PrYour Research Poster a

Your poster will be exposed to ov• delegatesReceive $50 off your registration: •$50 discount you must submit youposter title when registering for theYour poster abstract will be publish•conference CDYour research will be seen by lead•pharmaceutical, biotech, academicment institutes

To secure a poster board and inclusconference CD, your abstract must accepted,and registration paid in ful2009. You will be prompted to submabstract electronically when you regmeeting.

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PEGSummit.com15

Second Annual

Immunogenicity of Therapeutic BiologicsBioanalytical Assays, Preclinical and Clinical Assessment April 8-9

Recommended Short Course*SUNDAY, APRIL 5

1:00pm Short Course Registration2:00 – 5:00 pm (SC6) Testing for ImmunogenicityDifferent types of bioanalytical assays need to be developed and validated to support the development of therapeutic proteins. This short course will discuss issues encountered with the development and validation of bioanalytical assays as well as planning and interpreting immunogenicity testing in preclinical and clinical development programs.Instructors:David W. Scott, Ph.D., Professor of Surgery, Microbiology/Immunology, University of Maryland, School of Medicine, Center for Vascular and Infl ammatory DiseasesMatthew Baker Ph.D., Chief Scientifi c Offi cer, Antitope Ltd Harald Kropshofer, Ph.D., Global Coordinator Immunosafety, F. Hoffmann La Roche AG Mauricio Maia, Ph.D., Scientist, Genentech

*Separate Registration RequiredMAIN CONFERENCE

WEDNESDAY, APRIL 87:30am Registration and Morning Coffee

ASSESSMENT AND DEVELOPMENT OF ASSAYS FOR IMMUNOGENICITY

Screening• Confi rmatory Assays• Strategies to Address Drug Interference•

8:30 Chairperson’s Opening RemarksLawrence N. Callahan, Ph.D., Chemist, Offi ce of the Commissioner, FDA


8:40 Immunogenicity Assessment: Things to Consider When Developing the Strategic Plan

Steven J. Swanson, Ph.D., Executive Director, Clinical Immunology, Amgen Inc.The fi eld of immunogenicity assessment continues to evolve as new methods are developed and new expectations from regulatory agencies are developed. When deciding on the strategy to assess the immunogenicity of new protein therapeutics there are several important factors that should be considered. This presentation will highlight some of those important factors.

9:10 Next Generation Biologics: Deimmunization and Tolerance Induction

Anne S. De Groot, M.D., CEO/CSO EpiVax, Inc.; Professor and Director, Institute for Immunology and Informatics, University of Rhode Island; Pediatric Infectious Disease, Brown University Medical SchoolWe have developed and validated a set of tools with three primary applications to drug development: 1) ranking of the overall immunogenicity of proteins according to their puta-tive T-cell epitope content, 2) analysis of the protein for clusters of T-cell epitopes (the underlying cause of immunogenicity), and 3) pinpointing of key residues that can be stra-tegically altered to disrupt those epitope clusters. In addition, we have identifi ed a set of putative natural T regulatory epitopes (Tregitopes) which, when co-administered with an antigen, cause the expansion of antigen-specifi c adaptive. We have now confi rmed that co-administration of Tregitopes with a range of proteins (such as Ovalbumin, Botulinum toxin peptides, Dust mite antigen, fl u epitopes) in vitro and in vivo leads to suppression of T cell and antibody responses to the test antigens. The mechanism of suppression ap-pears to be due to the induction of antigen-specifi c adaptive tolerance induction (De Groot AS et al. Activation of natural regulatory T cells by IgG Fc-derived Peptide “Tregitopes” Blood (2008) 112: 3303).

9:40 Immunogenicity Assay Cut-point(s)Eric Wakshull, Ph.D., Senior Scientist & Group Leader, Bioanalytical R&D, Genentech, Inc.Recent guidance white papers have recommended a risk-based approach to developing immunogenicity strategies and assays in support of protein therapeutic pre-clinical and clinical development. The current recommended approach for establishing a cut-point for screening assays based upon population variability and a target 5% untreated posi-tive rate for drug-naïve samples has been extended to include methods for determining cut-points for confi rmatory and titration assays. The basis for unifi ed approach will be de-

scribed for all three cut-points, and examples provided. We feel this consistent approach provides an unbiased cut-point at each step of sample analysis and in particular helps solve the common observation of titer samples not crossing their cutpoint.

10:10 Coffee Break in the Exhibit Hall11:10 Immunogenicity and T cell responses: What Can We Learn

from Monitoring T-Cell Responses to Autoantigens?Vicki Seyfert Margolis, Ph.D., Chief Scientifi c Offi cer, Immune Tolerance Network Due to very low frequencies of autoantigen-specifi c T cells, their detection may be prob-lematic and requires very sensitive methods for monitoring various cell functions. We developed and optimized conditions for an Elispot assay that enables us to monitor re-sponses to autoantigens in Type 1 Diabetes and Multiple Sclerosis. This assay will be introduced for monitoring T-cell immunogenicity responses.

11:40 Adverse Reactions to Biologics: Accurate and Quantitative Measurement of Specifi c Antibodies

Jörgen Dahlström, Ph.D., Phadia ABPhadia has 40 years of experience in quantitative measurement of specifi c antibodies. With a yearly production of 70 million tests Phadias ImmunoCAPTM technology has a proven track-record with high sensitivity, user independent results, regulatory approval and long term stability making it well suited for measuring specifi c antibodies to biologics in clinical development.

12:10 Immunogenicity and Cell Based Sponsored by Assays on the Meso Scale Discovery (MSD) PlatformNeeta Shenoy, Ph.D., Scientist, Meso Scale DiscoveryMeso Scale Discovery offers signifi cant advantages for the rapid development and robust implementation of immunogenicity assays in preclinical and clinical settings to detect and characterize antibodies produced in response to biological therapeutics. High sensitivity, broad dynamic range and tolerance to free drug, along with reduced matrix effects and a simplifi ed workfl ow renders the platform ideally suited to address the challenges of im-munogenicity assays. MSD also provides solutions for cell based assays including NAb (neutralizing antibody) assays. Cell based assays on the MSD platform can range from quantitation of secreted proteins such as cytokines, monitoring changes in receptor phos-phorylation status or modulation of one or more intracellular markers, to characterizing interactions between proteins and cell surface receptors.

12:40 Luncheon Presentation II (Opportunity Available)1:10 Break1:30 Risk Assessment of Immunogenicity and Clinical ExpectationsDeborah Finco-Kent, Ph.D., Immunogenicity Lead, Drug Safety Research & Development, Pfi zer Inc. This talk will provide some representative nonclinical and clinical case studies with several types of biological therapeutics. Each case study will include a discussion regarding risk assessment as it relates to immunogenicity, Immunogenicity assessment plans for the particular program and regulatory feedback with respect to immunogenicity assessment.

2:00 Problem Solving Break-out Session: Drafting a Risk-based Immunogenicity Plan for a New Biologic Candidate

Facilitators:

Table 5Darshana Jani, Senior Associate Scientist, Biogen Idec Inc and Jaya Goyal, Associate Director, Clinical Science & Technology, Biogen Idec Inc.

Table 6Eric Wakshull, Ph.D., Senior Scientist & Group Leader, Bioanalytical R&D, Genentech, Inc.

Table 7Bonita Rup, Assistant Vice President, Protein Bioanalysis, Bioanalytical R&D, Drug Safety and Metabolism, Wyeth Research

Table 8Deborah Finco-Kent, Ph.D., Immunogenicity Lead, Drug Safety Research & Development, Pfi zer Inc.


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PRECLINICAL ASSESSMENT OF IMMUNOGENICITY3:45 Chairperson’s RemarksJoy Cavagnaro, Ph.D., DABT, RAC, President, Access BIO LC

3:50 The Value of Immunogenicity Assessment in Preclinical DevelopmentJoy Cavagnaro, Ph.D., DABT, RAC, President, Access BIO LCSince administration of human proteins to animals is expected to eliciting an immunological response the primary objective of assessing preclinical immunogenicity is to better defi ne the PK/PD profi le, exposure margins and estimates of toxicity which may be impacted by antidrug antibodies. Animal models can also be useful in assessing relative immunogenicity and in validating de-immunization strategies.

4:20 A Regulatory PerspectiveLawrence N. Callahan, Ph.D., Chemist, Offi ce of the Commissioner, FDA

4:50 Networking Cocktail Reception in the Exhibit Hall6:00 Close of Day

THURSDAY, APRIL 9

CLINICAL IMPACT OF IMMUNOGENICITY How do you evaluate immunogenicity in the clinic? • What does it mean? • What do you do about it?•

8:30am Chairperson’s RemarksMichel Awwad, Ph.D., Director, Bioanalytical R&D, Wyeth Research

8:35 Case StudyBonita Rup, Assistant Vice President, Protein Bioanalysis, Bioanalytical R&D, Drug Safety and Metabolism, Wyeth ResearchAssessment of immunogenicity risk and development of a testing strategy begins in the early nonclinical evaluation of a biotherapeutic protein product but should continue to evolve throughout the life cycle of a product. This presentation will overview the rationale and outcome of the proactive and reactive evolution of the testing strategy for a therapeutic protein product as it moved from fi rst in human studies into clinical trials and eventually into post-marketing study and monitoring stages.

9:05 Case Study: Clinical Impact of Immunogenicity and Implications Down StreamLakshmi Amaravadi, Ph.D., Associate Director, Preclinical & Clinical Development Sciences, BiogenIdecIn this presentation, case study examples to illustrate strategies implemented to evaluate immunogenicity will be presented. Using the example, clinical impact of immunogenicity on safety and effi cacy and correlation to clinical end points will be discussed. Approaches used in classifying the type of immune response in patients depending on clinical impact will be reviewed. Presentation will also discuss implications of immunogenicity data beyond product approval and discuss considerations pharmacovigilance approaches that are appropriate depending on the drug product.

9:35 Clinical Experiences on Impact of Immunogenicity on Safety and Effi cacy of Protein TherapeuticsNarendra Chirmule, Ph.D., Executive Director, Clinical Immunology, AmgenAll protein therapeutics have the potential of generating an immune response. Since this immunogenicity has the likelihood of impacting safety and effi cacy, regulatory agencies require extensive immunogenicity evaluation for these products. The immunogenicity testing strategy is driven by a risk-based approach. The clinical impact of immunogenicity on safety, ef-fi cacy, and the importance of long-term follow-up will be discussed with case studies.

10:05 Coffee Break in the Exhibit Hall11:05 Adnectins - Case Study from the Clinic of the Impact of ImmunogenicityEric Furfi ne, Ph.D., Senior Vice President, Research & Preclinical Development, Adnexus, a Bristol-Myers Squibb R&D CompanyAdnectins are a novel, proprietary class of targeted biologics that are derived from human fi bronectin, which is a well-characterized, highly-expressed plasma protein. Adnectin-based products offer potential advantages compared to traditional protein therapeutics, including speed of discovery, ease of manufacturing, and the ability to create an array of multi-functional targeted biologics. CT-322, an inhibitor of VEGFR-2, is the lead Adnectin now in phase II clinical development for the treatment of glioblastoma. Emerging clinical data suggests a low-risk immunogenicity profi le for this new Adnectin product. In addition, Adnexus has used its PROfusion discovery engine and EpiVax in silico technology to identify an early-stage, potent Adnectin with low immunogenic potential that addresses an important cancer target. Results of these programs will be discussed as case examples of how one can successfully manage or improve immunogenicity risks within a new biologics class.

11:35 Actual Impact of Immunogenicity in Clinical Outcome: Clinical Data of Myozyme TreatmentSusan M. Richards, Ph.D, Group Vice President, Clinical Laboratory Sciences, Genzyme Corporation

12:05 End of Immunogenicity of Therapeutic Biologics conference

PEGSummit.com17

Sixth Annual

Monoclonal AntibodiesAdvancing Conventional Antibody Development and Production

April 9-10Recommended Short Course*

SUNDAY, APRIL 51:00pm Short Course Registration

2:00-6:00pm (SC4) Translational Strategies for Development of Monoclonal Antibodies from Discovery to the Clinic

The effective information fl ow and translation of accumulated knowledge across various antibody development stages remain a major challenge. Successful strategies for development of monoclonal antibodies require integration of relevant knowledge with respect to target antigen properties, antibody design criteria such as affi nity, isotype selection, pharmacokinetic (PK)-pharmacodynamic (PD) properties, and antibody cross-reactivity across species from the early stages of antibody development. Biophysical measurements are one of the critical components necessary for the design of effective translational strategies for lead selection and evaluation of the relevant animal species for preclinical safety and effi cacy studies. Incorporation of effective translational strategies from the early stages of the antibody development process is a necessity; when considered it not only reduces development time and cost, but also fosters implementation of rational decision making throughout all phases of antibody development.Instructors:Mohammad Tabrizi, Ph.D., Director, Global PK-PD and Bioanalysis, MedImmune, Hayward CAGadi Bornstein, Ph.D., Principal Scientist, AstraZeneca R&D BostonScott L. Klakamp PhD, Research Fellow, Biophysical Chemistry and ResearchInformatics, Takeda San Francisco

*Separate Registration is Required

12:00pm Conference Registration

ADVANCES IN BASIC SCIENCE1:30 Chairperson’s Opening Remarks1:40 Developmental Trends in Antibody Fragment TherapeuticsAaron Nelson, Ph.D., Tufts University School of MedicineMonoclonal antibody derived fragments are a nascent category of targeted therapeutic agents, and a seemingly endless array of molecules can be engineered from various antibody domains. We sought to determine trends in the development of 38 preclinical antibody fragments and 47 therapeutic molecules that have entered clinical study sponsored by commercial fi rms. Our fi ndings indicate antibody fragment therapy is an emerging technology that faces a major hurdle at the proof-of-clinical-concept stage. Development may shift from FABs to scFvs and third-generation molecules, and from oncology indications to immunologic diseases.

2:10 Extending the Plasma Half-life of Antibody Fragments and Other Biopharmaceutical Proteins via Genetic Fusion with Homo-amino-acid Polymer Sequences

Arne Skerra, Ph.D., Professor of Biological Chemistry, Technical University MunichGenetic fusion with conformationally disordered homo-amino-acid polymer sequences provides an alternative way to attach a solvated random chain with large hydrodynamic volume to the protein of biopharmaceutical interest. Such sequences confer high solubility, are resistant against serum proteases, and lead to a large apparent size of the fusion protein. Moreover, they permit effi cient production of biochemically active fusion proteins in E. coli. Investigation of the plasma half-life for correspondingly modifi ed antibody fragments and cytokines reveals a pronounced prolongation effect depending on the length and composition of the polymer chain. In particular, genetic fusion with sequences comprising Pro, Ala, and Ser residues can yield pharmacokinetic properties similar to PEGylation, yet without necessitating costly and laborious in vitro modifi cation steps. This PASylation strategy should be useful for the development of therapeutic single-chain and single-domain antibody fragments.

2:40 Quartz Crystal Microbalance Biosensors in Immunology Anti-Malarial Antibody Affi nity/Avidity Issues Sponsored by

Lars M. Jørgensen, Ph.D., University of Copenhagen, Institute for Medical Microbiology and ImmunologyMalaria is widespread in tropical and subtropical regions, including parts of the Americas, Asia, and Africa. Each year, there are approximately 515 million cases of malaria, killing between one and three million people; the majority of whom are young children. Malaria is commonly associated with poverty, while it also positively stimulates poverty to become a major hindrance to economic developments. Malaria has currently become one of the most common infectious diseases and an enormous public health problem. Although some vaccines are under development, there is currently none available for malaria. Current preventive drugs must be taken continuously to reduce the risk of infection. Here we use the Attana Quartz Crystal Microbalance technology, as an innovative tool to better understand the mechanism and to improve the development of anti-malarial antibodies.

3:10 Refreshment Break in the Exhibit Hall4:00 Critical Role of Somatic Mutations in High Potency Human

Monoclonal Antibodies to VirusesJames Crowe, M.D., Ingram Professor, Pediatrics and Microbiologyand Immunology, Vanderbilt University Medical CenterOur laboratory has developed novel approaches to development of human monoclonal antibodies that rapidly yield high potency antibodies against viruses. We have isolated potent human monoclonal antibodies from human B cells directed to major human pathogens, including RSV, 1918 and H5 avian infl uenza, rotavirus, HIV and others. Genetic, biochemical and structural studies reveal that the principal driving force for optimal development of neutralizing human monoclonal antibodies is somatic hypermutation.

4:30 A Novel HPLC-UV-MS Method for Quantitative Analysis of Protein Glycosylation

Anton S. Karnoup, Ph.D., Analytical Sciences, The Dow Chemical Company

5:00 Close of DayFriday, April 10

ACCELERATING THE PROCESS7:45 Continental Breakfast in the Exhibit Hall 8:30am Chairperson’s Remarks


8:35 Two-Headed Antibody-Like Molecule: The Future of Cancer Antibody Therapeutics

Matthew Robinson, Ph.D., Associate Member, Fox Chase; Researcher, Fox Chase Head and Neck Cancer Keystone Program Bispecifi c antibodies that co-target two distinct tumor associated antigens, such as ErbB2 and ErbB3, can be an effective approach to disrupt critical signaling pathways relevant to tumor biology. Bispecifi c antibodies can have the added benefi t of providing enhanced tumor targeting selectivity; a property that is potentially benefi cial for the development of new immunodrug conjugates.

9:05 Selection of Antibody Producing Cells by Toxine ConjugatesKatrin Messerschmidt, Ph.D., Biotechnology, University of PotsdamThe generation of antibodies with designated specifi city comprises cost-intensive and time-consuming screening procedures. Here we present a new method by which hybridoma cells can be selected based on the specifi city of the produced antibody by the use of antigen-toxin-conjugates thus eliminating the need of a screening procedure. Initial experiments were done with several low molecular weight toxins and fl uorescein as model antigen. Several fl uorescein-toxin-conjugates were generated, purifi ed by HPLC and characterized regarding their toxicity.First results show that hybridoma cells that produce fl uorescein specifi c antibodies are able to dispose fl uorescein-toxin-conjugates.

9:35 Lead Identifi cation and Optimization in Crude Samples using Label-free Resonant Acoustic Profi ling

Helge Schnerr, Ph.D., Product Manager, TTP LabTech, TTP LabTech Ltd.Although optical-based systems dominate the biosensor market, piezoelectric and acoustic devices represent similar but signifi cantly less expensive alternatives. Acoustic biosensors have been employed in the label-free detection of an incredibly broad range of analytes; from interfacial chemistries and lipid membranes to small molecules and whole cells. Resonant Acoustic Profi ling (RAP) technology offers label-free, real-time analysis of biomolecular interactions. The analytical capabilities of a RAP-based biosensor are ideally suited to the development of biotherapeutics, a rapidly expanding area of drug research. In particular, direct measurement in crude and complex samples such as cell culture media or periplasmic extracts eliminates expensive time-consuming purifi cation of often limited material while delivering high content information.


PEGSummit.com 18

11:05 Cooperation of Dendritic Cells with Naive Lymphocytes on Artifi cial Extracellular Matrix to Accomplish in vitro Stimulation of Immune Response

Burkhard Micheel, Ph.D., Biotechnology, University of PotsdamTo serve as a basis for an in vitro immunization a cell culture model was established which combines an artifi cial extracellular matrix of the paracortical area of lymph nodes with involved immune cell populations of Ova-TCR-transgenic mice and defi ned antigen. The interaction of dendritic cells, T cells and B cells was investigated by morphological studies, immunfl uorescence staining and secretion of interleukins and antibodies. Ovalbumin-specifi c antibodies could be detected in the culture supernatants after co-cultivation of naive B cells with T cells and dendritic cells. These studies indicate, therefore, that modifi cations of cell culture substrata with extracellular matrix components support a stronger in vitro interaction of immune cells than plain surfaces and that this effect can be used for the induction of the production of specifi c antibodies in vitro.

11:35 Pre-Clinical Selection of Non-Immunogenic Lead Antibodies by the Accurate Detection of T Cell Epitopes

Matthew Baker, Ph.D, CSO, Antitope, Ltd.Helper (CD4+) T cell epitopes are drivers of immunogenicity through induction of class-switched, somatically mutated high affi nity antibodies which can bind to and neutralize the injected therapeutic. Various methods have been developed to detect T cell epitopes in the sequence of therapeutic antibodies. Several groups have claimed correlation of high affi nity MHC class II binding peptides (pMHC) detected using in silico methods with the presence of T cell epitopes, and have proposed the removal of such pMHC. Data will be presented to demonstrate that this approach is fl awed as some high affi nity pMHC can induce immunological tolerance, whilst certain low affi nity pMHC result in effector CD4+ T cell responses that can mediate unwanted immune reactions.


CASE STUDES1:25 Chairperson’s Remarks1:30 Development and Characterization of Monoclonal Antibodies

to Equine Luteinizing Hormone for Diagnostic ApplicationsNathalie Forster, M.S., Product Development and Support Manager, Research and Development, Maine Biotechnology Services, Inc.The success of a hybridoma antibody development project is contingent upon many factors. Among these factors include the integrity of immunizing and screening reagents, the ability of the fusion screening strategy to discern fusion products of interest, and the capability to characterize the resulting antibodies for use in the end application. In an effort to generate monoclonal antibodies to equine luteinizing hormone(eLH) suitable for the veterinary reproductive diagnostic market, MBS recently embarked on a hybridoma development project. Data from this hybridoma project will be presented as well as insight into the development strategies that lead to the successful production of several high affi nity, well-characterized monoclonal antibodies.

2:00 High Cytotoxic mAb for the Treatment of FMAIMargarita Salcedo, Ph.D. Pharmacology & Toxicology Department Director, LFBThe LFB has developed a proprietary antibody production process leading to the generation of monoclonal antibodies with high ADCC activity and enhanced affi nity for CD16, correlated to its glycosylation pattern. A fi rst generation anti-RhD candidate (R297) has been studied in human healthy volunteers. This monoclonal has been the fi rst monoclonal antibody shown to clear RhD+ red blood as effi ciently as polyclonal anti-RhD antibodies in humans. The new generation anti RhD (LFB-593)product will enter into phase I clinical trial in Q4 2008.

2:30 Networking Refreshment Break3:00 Generation of Antibodies Against ToxinsDiana Pauly, Ph.D., Post Doctoral Researcher, Center for Biological Safety, Robert Koch-InstituteThe task of raising antibodies against toxins is fraught with one major obstacle: the toxicity of the toxins. In order to ensure survival of the immunized animals, several inactivation strategies have been used to address this problem. The traditional methods are time-consuming and to a variable degree labour-intensive. Therefore, we developed a novel immunization strategy: we covalently immobilized the native toxins onto microbeads in order to reduce their toxicity. We showed that ricin coupled to microbeads is at least 70 times less toxic than native protein in an in vitro cytotoxicity assay and the median lethal dose in mice was 12 times higher for the immobilized toxin than for the native protein. With this new technique we were able to generate highly specifi c and highly sensitive monoclonal and polyclonal antibodies against native toxins. The antibodies were either used for sandwich immunoassays, multiplex detection systems or functional blockade of the toxin.

3:30 Use of the ClonePix-FL in a Research Monoclonal FacilityRobin Barbour, BS, Associate Director Research, Biology, Elan PharmaceuticalsThe Genetix ClonePix FL™ is an automated fl uorescence-based colony selector that allows for high throughput screening of fusions and single step identifi cation of stable single cell clones. We have implemented the use of ClonePix Fl™ to increase our fusion productivity by approximately 50%, while decreasing our time from fusion to stable clone by 50%. The additional benefi ts of implementing this technology are rescue of previously lost clones, simple selection of high secreting clones and decreased potential of repetitive motion injury to our staff.

4:00 Targeting ED-B Fibronectin: Using BC1-IL12 (AS1409) in Melanoma and Renal Cell Carcinoma

Ben Doran, M.Sc., Research Scientist, Protein Production, Antisoma Research LimitedClinical responses have been observed to human interleukin-12 in phase I and Phase II trials but development of this molecule was abandoned due to systemic toxicity. It is hoped that targeting this activity by genetically fusing interleukin-12 to a humanised BC1 antibody binding the oncofetal splice variant of fi bronectin, extra domain B (ED-B), that reduced side effects and an increased therapeutic index can be observed. Immunohistochemistry shows ED-B to be expressed in renal cell carcinoma and metastatic melanoma which are tumors that have observed clinical effi cacy in previous trials involving naked interleukin-12, providing a clear rationale for development. An effi cient manufacturing process with high productivity, unusual for antibody fusion proteins, and high recovery has enabled Antisoma to initiate a phase I dose escalation study in these indications.

4:30 End of Monoclonal Antibodies Conference

PEGSummit.com 19

THURSDAY, APRIL 9

OVERCOMING PROTEIN CHALLENGES12:00pm Conference Registration1:30 Chairperson’s Opening Remarks


1:40 Amyloid Structure in Bacterial Inclusion BodiesSalvador Ventura, Ph.D., Associate Professor, Institute of Biotechnology, University of Barcelona Protein misfolding and aggregation have become extremely active areas of research in re-cent years. Of particular interest is the deposition of polypeptides into inclusion bodies in-side bacterial cells because it constitutes a major bottleneck in protein production, restrict-ing the spectrum of available protein-based drugs or three-dimensional protein structures. Now, we demonstrate unequivocally the existence of amyloid structures inside bacterial inclusion bodies. Moreover, we have been able to show that protein aggregation into inclusion bodies display an important specifi city, that the inclusion bodies are able to seed the formation of amyloid fi brils and the formation of SDS-stable oligomers of this peptide inside bacteria. These results suggest the existence of evolutionary conserved strategies to avoid the harmful effects of undesired protein aggregation by sequestering sticky folding intermediates into conformationally related stable aggregated structures through selective interactions in both eukaryotes and bacteria, highlighting that prokaryotic systems should be seriously considered when exploring the in vivo determinants and cellular effects of protein aggregation.

2:10 Endotoxin Removal from Biopharmaceuticals – A Challenge in Downstream Processing?

Stefan R. Schmidt, Ph.D., M.B.A., Associate Director, R&D, AstraZeneca ABLToday biopharmaceuticals are important therapeutics. For the manufacturing of protein drugs, various expression systems are used that can potentially be sources of endotoxins. One of the challenges in downstream processing is the reliable quantitative removal of endotoxins, and a number of technologies promise to solve this problem. This talk reviews the current approaches with regard to important parameters (e.g. effi ciency, cost, processing time, compatibility), de-scribes a novel scaleable technique, and gives recommendations for typical cases.

2:40 High Performance Simultaneous Clarifi cation and Capture Chromatography

Lisa Crossley, President & CEO, Natrix Separations


TECH TRANSFER4:00 A Case Study on Effective Strategies for Tech Transfer in

Biopharmaceutical ManufacturingRobert J. Broeze, Ph.D., President & CEO, Laureate Pharma, Inc., Princeton, NJ, and Thomas P. Loisel, Ph.D., Associate Director, Process Development, Enobia Pharma Inc.Tech transfer is an essential part of the development of every biopharmaceutical product as it moves from discovery to clinical and commercial production. Examples of effective strategies for effi cient tech transfer will be illustrated with a case study of a monoclonal antibody biopharma-ceutical. Transfer of technical know-how occurs throughout the development process from early developmental studies to process scale-up and transfer from client to CMO, through internal transfer from development and into cGMP production. Aspects of communication, interaction, facility design and logistics will be addressed.

4:30 Technology Transfer to Manufacturing: The Key to Successful Commercialization

Stephen M. Perry, President, Kymanox Inc.Success in protein manufacturing, and ultimately in the biotechnology business, is fundamentally linked to the technology hand-off to manufacturing. There are distinct differences between de-velopment and manufacturing groups and these differences must be managed actively to ensure transfer success. Furthermore, scale-up and transfer of protein processes have many potential pitfalls - some of which can be avoided when the right plan is in place. Lastly, technology transfer is a knowledge and communication activity where soft skills are as important as technical ones.

5:00 Close of Day

FRIDAY, APRIL 10

PROTEIN ANALYTICS7:45 Continental Breakfast in Exhibit Hall 8:30am Chairperson’s RemarksSalvador Ventura, Ph.D., Associate Professor, Institute of Biotechnology, University of Barcelona

8:35 PAT and Smart Chemometrics for Assured Pharma Development and Production

Julian Morris, Ph.D., Professor, Process Control, and Director, Centre for Process Analytics & Control Technology, Foresight Centre CPACT, Newcastle UniversityThe presentation will address the impact of ‘smart chemometrics’ in the production of high value-added products in the pharma-chemicals and bio-pharma industries through the introduction of PAT and QbD. With the increasing take-up of PAT, the issues related to the use of process analytical measurements for process understanding from product and process development, faster scale-up, consistent product output and assured downstream processing under closed loop process control depends heavily on developing robust calibrations using spectral measure-ments which are subject to fl uctuations in process variables, instruments and probes and in other external variables. A number of new chemometric algorithms capable of building robust ‘trans-ferable’ calibrations have recently been developed for FTIR, NIR, Raman Scattering and X-Ray diffraction (XRD) and these will be highlighted through application studies alongside highlighting the importance of moving PAT into closed loop process control.

9:05 Stability of Monoclonal Antibodies in Liquid FormulationsIvan Correia, Ph.D., Research Investigator, Protein Analytics, Abbott Bioresearch CenterMonoclonal antibodies (Mabs) are composed of an Fc region and two Fab regions linked by a fl exible hinge region. As a result of fl exibility, the hinge region is exposed and thus easily per-turbed and mabs in liquid formulation undergo non-enzymatic hydrolysis when stored at 5 C for extended periods of time. Hydrolysis in the hinge region can be enhanced at extreme pH and at high temperatures. In this presentation we identify a novel mechanism of cleavage in the hinge region of mabs.

9:35 Development, Manufacture, Preclinical Evaluation and Effi cacy Studies of a Melanin Binding IgM Antibody Labeled with 188Re Against Experimental Human Metastatic Melanoma in Nude Mice

Muctarr Sesay, Ph.D., Vice President, Process Development, Goodwin Biotechnology Inc.IgM monoclonal antibodies possess great challenges during their development and manufacture for human cancer therapy. This includes low cell culture productivity, limited purifi cation options, radiolabeling (of intact IgM) and characterization. This presentation will focus on an ongoing project and will describe strategies involving cell culture production, development and manu-facture of a purifi cation process that does not require the costly and ineffi cient use of Protein A affi nity column (most IgMs antibodies do not bind to this column) and resulted in therapeutic grade antibody. A fast, simple, cost effective and effi cient conjugation/modifi cation of the purifi ed IgM monoclonal antibody and subsequent radiolabeling of the modifi ed antibody with 188Re will also be presented.

10:05 Coffee Break in the Exhibit Hall11:05 The New Draft FDA Guidance on Process Validation- What

Does it Mean for You?James Blackwell, Ph.D., M.B.A., Senior Consultant, BioProcess Technology Consultants, Inc.The FDA issued new draft guidance to industry on process validation for comment in November of 2008. While not fi nalized, this guidance along with other “21st Century” initiatives, such as QbD, provide a road map that organizations can use now to approach their development efforts, whether in-house or outsourced. A comprehensive review of these initiatives and impact to key industry players will be buttressed with practical insights that can be used for the entire product development lifecycle.

11:35 A Modifi ed Manufacturing Process of Alpha-1 Proteinase Inhibitor Leading to Improved Product Yield and Purity

James Rebbeor, Ph.D., Purifi cation Science Manager, R&D & Technology, Talecris Biotherapeutics Inc.Our objective was to develop a high-purity intravenous alpha-1 proteinase inhibitor (PI) (TAL6004), for the treatment of congenital alpha-1 antitrypsin defi ciency as a line extension of the currently marketed Prolastin®. TAL6004 and Prolastin® production processes are based on Cohn VI plasma fractionation. The modifi ed process incorporates an enveloped virus inactivation step prior to anion exchange chromatography and a diafi ltration step after ultrafi ltration for added purity. Whereas the Prolastin® process employs a pasteurization step, the modifi ed process uses cation exchange chromatography and nanofi ltration for further purifi cation and pathogen removal. The fi nal bulk is ready after terminal ultrafi ltration/diafi ltration, sterile fi ltration and lyo-philization steps. The alpha-1 PI manufacturing process has resulted in increased purity and

Second Annual

Protein Scale-Up & ManufacturingHarnessing Complexity

April 9-10

PEGSummit.com20

yield of TAL6004 compared with Prolastin®. Extended characterization supports the process change with the two products having the same glycoform pattern as plasma alpha-1 PI. Scal-ability has been demonstrated from bench through commercial scale.


SCALE-UP CHALLENGES1:25 Chairperson’s RemarksJie Chen, M.D., Director, Protein Science, DYAX CORP.

1:30 Analytical Challenges to Establishing Comparability for Biologics: Raising the Regulatory Bar

Asenath Rasmussen, Ph.D., Associate Research Fellow, Analytical Research & Development, Pfi zer Worldwide Pharmaceutical Sciences Over the course of a product life cycle, modifi cations to the manufacturing process are almost inevitable. In the case of biologics, which are inherently heterogeneous, even minor changes to the process can impact the product profi le. As new analytical technologies are introduced, estab-lishing comparability becomes all the more challenging. This presentation will highlight specifi c analytical challenges and propose strategies that may be employed to address them.

2:00 Antibody Manufacture Pipelines - Optimized for Speed & QualityRalf Ostendorp, Ph.D., Senior Director R&D, Head of Protein Sciences, MorphoSys AG

2:30 Networking Refreshment Break

PROTEIN MANUFACTURING3:00 From Development to Large-Scale Manufacturing of

Immunoglobulins from Human PlasmaWolfgang Teschner, Ph.D., Director, Research, Plasma Product Development / Product Support Global Pre-clinical R&D, Baxter Innovations GmbHContinous growth of applications of human polyvalent immunoglobulin preparations are expect-ed in the next decade. In order to meet demand each year tons of immunglobulins are purifi ed out of human plasma. Purifi cation processes have to combine robustness in large-scale manufactur-

ing and adequate virus reduction of all types of viruses with a maximum IgG yield and purity. The fi nal product has to be easy to apply, effi cacious and well tolerated. Unchanged process parameters from pre-clinical to routine lot manufacturing assure the comparability of the results from each development stage.

3:30 Assessing the Strategic Value of Fast, Flexible Clinical Manufacturing Capacity

Geoffrey Hodge, Ph.D., Vice President, Process Development & Technology, Xcellerex LLC Biotherapeutics increase signifi cantly in value as they advance through clinical trials and ulti-mately to market. Clinical manufacturing facilities are large capital investments that must be planned many years in advance. Biotherapeutic development companies face the diffi cult chal-lenge of managing their pipeline in order to keep these strategic assets running at high capacity, but the uncertainties of drug development can cause “boom or bust” scenarios where plants sit idle or are unable to keep pace with clinical manufacturing demands. This talk will look at the economic modeling of several scenarios to determine the value of adding clinical manufacturing capacity as well as the value of being able to bring capacity online more quickly.

4:00 BioSMB: A New Continuous, Disposable Chromatography Process

Thomas Ransohoff, Ph.D., Consultant, Tarpon Biosystems Inc. & Vice President and Senior Consultant, BioProcess Technology Consultants Inc.The downstream processing bottleneck is leading to the exploration of processes with higher effi ciencies and higher productivities. BioSMB is a technology that refi nes traditional SMB into a viable option for biopharmaceutical purifi cation. By implementing a fully disposable-format fl uid path and modular design, the BioSMB technology addresses the key issues associated with biopharmaceutical processing, such as elimination of cleaning validation and rapid campaign change-over. This talk will present experimental data of a MAb downstream process using a bench-scale BioSMB system. The presentation covers the capture of MAb from cell supernatant on various commercially available Protein A media, packed in disposable cartridges. The data showed that the BioSMB outperformed the batch process in terms of MAb recovery and HCP reduction, while offering a signifi cant reduction in buffer consumption and a three to tenfold in-crease in productivity. Based on the experimental data, projections will be given for the impact of BioSMB technology on large-scale manufacturing of monoclonal antibodies.

4:30 End of Protein Scale-Up & Manufacturing Conference

THURSDAY, APRIL 912:00 pm Conference Registration

THE HOW AND WHY OF PROTEIN AGGREGATION - TOOLS, INTERPRETATIONS, AND CORRELATIONS1:30 Chairperson’s Opening RemarksMary Cromwell, Ph.D., Associate Director and Senior Scientist, Protein Analytical Chemistry, Genentech

1:40 Diversity and Complexity of Protein AggregationTudor Arvinte, Ph.D., Professor, University of Geneva; Chairman and Chief Executive Offi cer, Therapeomic, Inc.Development of biopharmaceuticals is often confronted with various types of protein aggregation phenomena. A complex battery of analytical method- including less distractive methods for loose aggregates- is needed to detect and characterize these aggregates. This talk would feature new and unpublished data to share different case studies that documents the diversity and complexity of protein aggregation phenomena.Detection and Quantitative Characterization of Aggregates

2:10 Detection and Quantitative Characterization of Protein-Protein Interactions in Highly Concentrated Solution

Allen P. Minton, Ph.D., Chief, Section on Physical Biochemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health

How protein – protein interactions in highly concentrated solution affect the • thermodynamic and colloidal stability of proteins and the physical properties of the solution How to characterize protein-protein interactions in highly concentrated solution via • measurement of static light scatteringHow to characterize protein-protein interactions in highly concentrated solution via • measurement of tracer sedimentation equilibrium

2:40 Polyglutamine Domains as Novel Intra-Molecular AdjuvantsAlexander Shneider, Ph.D., Founder and CEO, Cure Lab, Inc.Alum and oil droplets have been used as successful vaccine adjuvants due to their ability to absorb antigenic proteins on their surface, thus creating antigen-containing macroparticles. However these adjuvants can not be applied to recombinant vaccines against infectious diseases and cancer. Contrary to currently used subunit and/or inactivated vaccines, a recombinant vaccine is a vector expressing an antigenic protein within a cell. Intracellular aggregation of an antigenic protein (preferably, followed by the protein release from the aggregates) would act similarly to alum adjuvant. Therefore, a new era in vaccinology requires novel molecular adjuvants inducing aggregation of antigenic proteins within a transected cell. Here we report that aggregate-forming polyQ domain fused to an antigenic protein serves as an intra-molecular adjuvant augmenting both B- and T-cell immunity.

Gain information about a new adjuvant, which can be used by an interested party • immediatelyLearn a novel approach to adjuvant development based on intracellular aggregation • of antigenic proteinsHear a crosstalk between two remote areas of biomedical science, vaccine • development and neurodegenerative disordersUnderstand a technology for effi cient expression of biologically and pharmaceutically • important cytotoxic proteins


Inaugural

Aggregation in Protein TherapeuticsUnderstanding and Overcoming Analytic, Formulation,Manufacturing, and Regulatory Challenges April 9-10

“ I now have a bett er insight into completely diff erent strategies that could solve various issues.”Section Leader, GlaxoSmithKline

PEGSummit.com 21

ANALYSIS OF AGGREGATES AND SUB-VISIBLE PARTICLES IN PROTEIN SOLUTIONS4:00 Detecting Aggregates in Protein Therapeutics without Sample

Separation or Dilution by Low-angle Dynamic Light ScatteringRoland Schmidt, Ph.D., Principal Scientist, Manufacturing Science and Technology, Abbott LaboratoriesWhen applying the traditional array of analytics for the detection of protein aggregates, the analyst is often confronted with the question of the impact of sample dilution and separation on protein aggregate structure and detectability. Only few technologies are available with the capability to analyze aggregates in high-concentrated protein solution without sample preparation. Data will be presented highlighting low-angle dynamic light scattering as promising tool for analyzing high-concentrated protein solutions for higher-order aggregates and particulate matter in undiluted state.

Analytical challenges associated with high-concentrated protein solutions• Dynamic light scattering technology and applications for protein therapeutics• Low-angle dynamic light scattering capabilities and limitations•

4:30 Analysis of Aggregates and Sub-visible Particles in Protein Solutions: Case Studies

Yijia Jiang, Ph.D., Principal Scientist, Formulation and Analytical Resources Department, AmgenAggregation and particle formation have become the focus of increased attention for the biopharmaceutical industry recently due to increasing concerns about their impact on the safety and effi cacy of biological products. Aggregates and particles span the size range from a few nanometers to hundreds of micrometers. Little actual data on their impact on product quality is available. The fi rst step in obtaining data on the safety impact of these species is to be able to reliably analyze them. Both the characterization of these particles, and understanding the underlying mechanism of their formation, present challenges. Case studies showing the analysis of aggregates and sub-visible particles will be presented.

5:00 Close of Day5:00 - 5:30 Registration for Aggregation Dinner Short Course

Presenter and Panel Moderator: John Philo, Ph.D., Vice President, Biophysical Chemistry, Alliance Protein LaboratoriesThe introductory talk will review the types and mechanisms of aggregation, and why characterization methods in addition to SEC are often requested by the regulators. Advanced methods for characterization of aggregation will be described, including analytical ultracentrifugation (AUC) and light scattering techniques (SEC-MALS and DLS). The advantages and disadvantages of these techniques will be discussed, along with some real-world examples and applications. The audience will gain a better understanding of mechanisms of aggregation, how those mechanisms point to prevention, and which analytical methods should be chosen. Thereafter, the audience and panel members will discuss the issues and learn from each others’ challenges. The presentation and interactive panel discussion will cover:

Types and mechanisms of aggregation• Why characterization methods in addition to SEC are often requested by the regulators• Basic principles of orthogonal analytical methods analytical ultracentrifugation (AUC), • fi eld-fl ow fractionation (FFF) and light scattering techniques (SEC-MALS and DLS)Advantages and disadvantages of all these analytical methods• Real-world examples and applications to illustrate a range of aggregation issues and • mechanisms that may be encountered and how these various analytical methods can be applied

Panelists:Yijia Jiang, Ph.D., Principal Scientist, Formulation and Analytical Resources Department, AmgenTudor Arvinte, Ph.D., Professor, University of Geneva; Chairman and Chief Executive Offi cer, Therapeomic, Inc.Mary Cromwell, Ph.D., Scientist & Senior Group Leader, Early Stage Pharmaceutical Development, GenentechAbout the Guest Presenter and Panel Moderator:John is the V.P. and co-founder of Alliance Protein Laboratories, where he is in the laboratory analyzing aggregation nearly every day for more than 150 clients world-wide. Before co-founding A.P.L. in 1998 John spent over 6 years in Protein Chemistry at Amgen, where his primary responsibilities were protein characterization using analytical ultracentrifugation, light scattering, and calorimetry. While there he pioneered new sedimentation velocity techniques appropriate for biotechnology products and new software techniques for analysis of protein-receptor interactions by sedimentation equilibrium. Prior to working at Amgen, John was a faculty member in Molecular and Cell Biology at the University of Connecticut in Storrs for 9 years studying protein structure and function. While at U. Conn. he founded and headed the Macromolecular Characterization Facility in the Biotechnology Center and was a co-founder of the National Analytical Ultracentrifugation Facility. John was also a postdoctoral fellow at U. Conn. with Todd Schuster studying hemoglobin kinetics. He received his Ph.D. in Physics from Stanford in 1977. John has authored over 75 scientifi c publications including over 60 directly related to biotechnology and pharmaceutical applications.

(SC5) Dinner Presentation and Interactive Panel Discussion*Focus on Aggregation: Mechanisms, Analytical Methods,

and Real-World Examples 5:30 - 8:30 pm

At the close of the day an optional Dinner and Short Course will be hosted at the conference venue. Attendees must register in advance, as seating is limited. This event will feature both a talk and an interactive panel discussion where the audience can bring up specifi c issues or questions important to their work. * Separate registration is required

FRIDAY, APRIL 10

THE IMPACT OF PRODUCTION ON PROTEIN AGGREGATION7:45 am Continental Breakfast in the Exhibit Hall 8:30am Chairperson’s RemarksSambit R. Kar, Ph.D., Principal Scientist, Pharmaceutical R&D, Pfi zer Global Biologics

8:35 Opening Talk – Bioproduction: Many Sources of Protein AggregationMary Cromwell, Ph.D., Associate Director and Senior Scientist, Protein Analytical Chemistry, Genentech

9:05 Infl uence of Formulation on the Success of Protein DrugsTudor Arvinte, Ph.D., Professor, University of Geneva; Chairman and Chief Executive Offi cer, Therapeomic, Inc.In spite of large efforts in biotechnology the number of new biotech products reaching the market each year is not high. Projects may die due to the use of not optimized protein formulations and the failure is falsely attributed to the molecule instead. The presence of protein aggregates may be linked to the side effects and lack of activity of protein drugs. The talk will be present case studies where small differences in formulation have unexpected consequences in the aggregation state of biopharmaceuticals . One challenge for protein formulation is to analyze protein aggregation in the formulation in conditions as near as possible to those in which the drug is applied in vivo. These “tailor-made” analytical methods should be adapted to the necessities of the formulation rather than to the needs of the analytical techniques.

9:35 Mechanical Stress Stability of Monoclonal Antibody Formulations

Sylvia Kiese, Ph.D., Formulation Scientist, Pharmaceutical R&D Biologics, F. Hoffmann-La Roche Ltd.Protein aggregation is known to occur under different stress conditions and displays a wide variety of morphologies and sizes. The detection of aggregate formation in terms of morphology, size and number requires various analytical techniques, each with their advantages and limitations. The focus of this presentation is to analyze and compare protein aggregation induced by various mechanical stress setups using an array of analytical methods.


CHARACTERIZING AGGREGATION TO IMPROVE PRODUCT YIELD AND THE PURIFICATION PROCESS11:05 Characterization of Soluble Aggregates in Formulation

DevelopmentSambit R. Kar, Ph.D., Principal Scientist, Pharmaceutical R&D, Pfi zer Global BiologicsThis presentation will discuss the following key points:

Soluble aggregates in biotherapeutics• Biophysical characterization• Analytical ultracentrifugation• Reversible aggregation•

11:35 A Case Study Detailing the Application of Analytical Methods to Characterize Protein Aggregation for a Biopharmaceutical Drug Program

Jennifer F. Nemeth-Seay, Ph.D., Principal Research Scientist, Head, Discovery Mass Spectrometry, Centocor Research and Development The expression and purifi cation of protein drug candidates is a complex process that does not produce just the expected product, but by-products as well. Often, drug impurities such as protein clips, multimers, or aggregate, are also produced. In order to fully understand the composition of a protein product, which could be a combination of all the above species; as well as to improve future production methods; it is important to characterize these other protein forms during the course of development. In this case study, a variety of analytical techniques including SDS-PAGE gels, size-exclusion chromatography (SEC), SEC-static light scattering (SLS), free-thiol analyses, and mass spectrometry were used to characterize aggregate found in multiple lots of material of a biopharmaceutical drug candidate. The information is being used to improve product yield and the purifi cation process.

12:05pm Luncheon Presentation I (Opportunity Available)12:35 Luncheon Presentation II (Opportunity Available)1:05 Coffee Break in the Exhibit Hall1:25 Chairperson’s RemarksAllen P. Minton, Ph.D., Chief, Section on Physical Biochemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health

1:30 Removal of IgG2 Aggregates by Hydrophobic Interaction Chromatography

Min Zhu, Ph.D., Scientist, Process Biochemistry, MedImmuneThis presentation describes the development of an aggregate removal step using hydrophobic interaction chromatography (HIC). For this study an IgG2 monoclonal

PEGSummit.com22

antibody that was prone to aggregation was subjected to biophysical characterization. Results of ANS binding and reverse phase chromatography suggested that the aggregate form was more hydrophobic than the native monomer. In addition, circular dichroism (CD) and peptide mapping analysis indicated that the aggregate exhibited a different tertiary structure and disulfi de linkage pattern compared to the native monomer. These differences suggested that HIC could be used to selectively separate the aggregate from the native monomer. The development of an HIC step is often challenging compared to other chromatography separation methods because of the number of factors that must be considered in optimization, including ligand and backbone of the chromatography gel, as well as buffer composition, pH, salt selection and conductivity. For this reason, a streamlined approach was undertaken that combined gel selection with the evaluation of the binding capacity in a high throughput screening format. This was followed by the development of step elution conditions designed to achieve a high resolution resulting in a simple and robust unit operation. This development strategy permitted the successful rapid design and optimization of a HIC chromatography step meeting requirements for aggregate removal within very tight project timelines. Although an alternative ion exchange step was also developed and optimized in parallel, the HIC step proved to be equal or superior with respect to product purity and yield.

OVERCOMING CHALLENGES TO PROTEIN FORMULATION AND AGGREGATION IN THE FROZEN STATE2:00 Avoiding Aggregation in Bulk Freeze and Thaw – How Much

and What Kind of Control you Need – A ControversyRoland Schmidt, Ph.D., Principal Scientist, Manufacturing Science and Technology, Abbott LaboratoriesFreezing and thawing of protein bulk solution for storage and shipping is a process routinely used in biologics manufacturing. Freezing in general is known to bear a potential for protein aggregation, which calls for a high level of control for all freeze and thaw unit operations. A controversy, however, exists in the fi eld about the practical implementation of the required process control. On the one hand, a traditional approach utilizing small individual containers in conventional freezers is employed by numerous marketed products. On the other hand, specialized technologies applying well-defi ned freeze and thaw rates are being implemented throughout the industry. Case studies for both approaches will be discussed highlighting justifi cation strategies utilizing Quality-by-Design concepts.

Freezing and protein aggregation• Current technologies for bulk freezing of biologics• Approaches to control freezing and thawing processes and avoid aggregation•

2:30 Networking Refreshment Break3:00 Protein Aggregation in the Frozen State due to Excipient

CrystallizationDeirdre Piedmonte, Ph.D., Senior. Scientist, Formulation and Analytical Resources, Amgen Inc.We recently identifi ed aggregation of low protein concentration formulations (stored at -30 °C) that occurred due to crystallization of the commonly used excipient sorbitol (Pharm Res. 2007 24(1):136-46). The cause of protein aggregation was phase separation of the protein and its stabilizing excipient that occurred during long-term frozen storage because the excipient crystallized. We have expandedthe scope of our previous work to evaluate the effect of formulation variables such as protein concentration and pH on the sorbitol crystallization-induced aggregation of monoclonal antibody formulations. We have identifi ed formulation conditions that prevent sorbitol crystallization-induced aggregation, which may then provide the fl exibility to freeze sorbitol-containing formulations. We also describe methods to identify formulation conditions where excipient crystallization may occur, providing a faster alternative to real-time studies.

By sharing our practical experience in overcoming challenges to protein formulation • in the frozen state, the audience will gain an understanding of a cause and potential prevention of protein aggregation.On a broader note, our presentation will raise awareness that the evaluation of • potential changes to excipients during product storage is essential to drug stability and product shelf life.Audience members will gain an appreciation of the importance of selecting excipients • (with respect to potential physical changes throughout storage) during formulation development and the importance of integrating excipient characterization into routine formulation development programs, particularly in light of increasing QbD expectations of regulatory Agencies.

BIOLOGICAL EFFECTS OF PROTEIN AGGREGATES3:30 The Role of Aggregates Inducing Antibody Responses Against

Protein TherapeuticsPhilippe Stas, Chief Executive Offi cer, AlgonomicsWith an ever increasing number of protein therapeutics reaching the patient, the specifi c challenges of this class of drugs are being characterized. Most protein therapeutics show immunogenicity, giving rise to anti-drug antibodies that in many cases are transient and harmless, but in some isolated cases lead to severe side effects. This presentation addresses the factors driving immunogenicity of protein therapeutic, with a specifi c emphasis on the role of aggregates inducing antibody responses against protein therapeutics. Strategies to identify and characterize immunogenicity are discussed in conjunction with novel regulatory guidance’s and industry whitepapers. Selected case

studies are presented.

4:00 AAA-ATPase p97/VCP: Cellular Functions, Disease and Therapeutic Potential

Neeraj Vij, M.S., Ph.D., Division of Pediatric Respiratory Sciences & Institute of NanoBioTechnology, The Johns Hopkins School of Medicinep97/VCP, dislodges the proteins from the ER and chaperones them for proteasomal degradation or aggregation. VCP has a polyQ and polyUb binding capacity and is involved in several neurodegenerative and misfolded protein diseases. We will discuss the therapeutic strategies to control VCP mediated protein aggregation or degradation in these diseases.Upon completion of this talk, participant should be able to:

Understand the mechanism of VCP mediated protein aggregation or degradation.• Identify the role of VCP in neurodegenerative and protein misfolding disorders• Learn novel therapeutic strategies and methods to target a specifi c protein function, • interaction or post-translational modifi cation.

4:30 End of Aggregation in Protein Therapeutics Conference

3M CoA&G Pharmaceutical Inc.A&G Precision AntibodyAbbott Bioresearch Center, Inc.Abbott LabsAbeome Corp.AblynxAbraxis ResearchAcceleron PharmaAccess BIOACS PublicationsAdAlta Pty, Ltd.Adimab, Inc.Adnexus TherapeuticsAffi body ABAffi tech ASAffymax, Inc.Aisling CapitalAjinomoto AminoScience LLCAlder iopharmaceuticalsAlexion Pharmaceuticals, Inc.AlgoNomics NVAllergan, Inc.Alligator BiosciencesAllozyne, Inc.Ambrx, Inc.Amgen, Inc.AmProteinAmunixAmylin Pharmaceuticals, Inc.Anaptys Biosciences, Inc.Apogenix AGApplied Molecular EvolutionAradigm Corp.Arana Therapeutics, Inc.Archemix Corp.ArecorArgonne National LabAscendis Pharma GmbHAssociates of Cape Cod, Inc.AstraZeneca ABLAstraZeneca Pharmaceuticals, Inc.Astrazeneca Pharmaceuticals LPAstraZeneca R&D MontrealAttana ABAuxilium Pharmaceuticals Inc.Avecia BiotechnologyAVEO PharmaceuticalsAvidBioticsAviGenicsAvila TherapeuticsBaxter HealthcareBaxter International, Inc.Bayer HealthCare PharmaceuticalsBayer Schering Pharma AGBD BiosciencesBeachhead ConsultingBend Research, Inc.Beth Israel HospitalBio Rad LabsBioAtlaBiocompare, Inc.Biogen Idec, Inc.BioInvent International ABBiologics Consulting GroupBioMarin Pharmaceuticals, Inc.Biomeasure, Inc.BioProcess Technology Consultants, Inc.Biotech Educational Network ServicesBioVectra DCLBioWa, Inc.Blue Sky Biotech, Inc.Boehringer Ingelheim Austria GmbHBoehringer Ingelheim Canada, Ltd.Boehringer Ingelheim PharmaBoehringer Ingelheim Pharma GmbH & CO KGBooz Allen HamiltonBristol Myers Squibb Co.Caliper Life Sciences, Inc.Caliper Technologies Corp.Cancer Research TechnologyCancer Research UKCatalyst BiosciencesCell Signaling Technology, Inc.Centocor R&D, Inc.Charles River Labs Preclinical Services Montreal, Inc.Charles Stark Draper Lab, Inc.ChemDiv, Inc.

Chesapeake Biological LabsChildrens Hospital BostonChugai Pharmaceutical Co., Ltd.Cisbio InternationalCleveland Clinic FoundationCNIOCNRSCoda GenomicsCodon Devices, Inc.Constant Systems, Inc.CovX Pharmaceuticals, Inc.CRTCSIRO AustraliaCSIRO Livestock IndustryCSIRO Molecular ScienceCSL, Ltd.CytomX LLCDaewoong Pharmaceuticals Co.Daiichi Pharmaceutical Co., Ltd.Daiichi Sankyo Co., Ltd.Dainippon Sumitomo PharmaDana Farber Cancer InstituteDaniscoDecision Biomarkers, Inc.DIREVO Biotech AGDoe & Ingalls of NC LLCDomantis, Ltd.Dow AgrosciencesDow Chemical Co.Dow Corning Corp.Dragonfl y Sciences, Inc.Dyax Corp.Dyax SAEcole Polytechnique Federale De LausanneEI DuPont De Nemours & Co.Eisai Co., Ltd.Elan PharmaceuticalsEli Lilly & Co.EMD Biosciences, Inc.EMD ChemicalsEMD Lexigen Research CenterEMD SeronoEmergent BioSolutionsEnobia PharmaEnzon, Inc.EpiVax, Inc.Ethicon, Inc.Eurofi ns MedinetExpert BioMed, Inc.ExSARF Hoffmann La Roche, Inc.FDA CDERFluidigm Corp.ForteBio, Inc.Fresenius Biotech GmbHFrommer Lawrence & Haug LLPFusion Antibodies, Ltd.Ganymed PharmaceuticalsGE HealthcareGedeon Richter, Ltd.GENEART AGGenelux Corp.Genentech, Inc.Genetic Engineering NewsGenetix, Ltd.Genetix USA, Inc.GENEWIZ, Inc.GENimmune BelgiumGenmab ASGenmab BVGenoFocusGenWay Biotech, Inc.Genzyme Corp.Gilead Sciences, Inc.GlaxoSmithKlineGlaxoSmithKline BiologicalsGlenmark PharmaceuticalsGlycArt Biotechnology AGGoodwin Biotechnology, Inc.Greenovation Biotech GmbHGWC TechnologiesHaematologic Technologies, Inc.Halozyme TherapeuticsHospira, Inc.Human Genome Sciences, Inc. HGSHyaluron, Inc.IBSM CNRSICx AgentaseICx NomadicsIdenix Pharmaceuticals, Inc.Idexx LabsIIBR

ImClone Systems, Inc.ImmunoGen, Inc.INETIInSight iopharmaceuticals, Ltd.IntrexonInvitrogen, Inc.Isogenica, Ltd.Johnson & Johnson Consumer Products, Inc.KEIKirin Pharma Co., Ltd.Kyowa Hakko Kirin Co., Ltd.Laureate Pharma LPLawrence Livermore National LabLilly VenturesLonza Biologics PlcLos Alamos National LabLucigen Corp.MacroGenics, Inc.Massachusetts General HospitalMayo ClinicMD Anderson Cancer CenterMedarex, Inc.MediGene AGMedImmune, Inc.MedImmune, Ltd.MedImmune R&D HaywardMerck & Co.Merck Research LabsMerialMerial, Ltd.Merrimack PharmaceuticalsMeso Scale DiscoveryMicrobiotix, Inc.MilleGenMillipore Corp.Mirus Bio Corp.Molecular Partners AGMonsantoMorgan & Claypool PublishingMorphoSys AGMorphotek, Inc.MSM Protein Technologies, Inc.MTF Musculosketal Transplant FoundationNational Research Council CanadaNature Publishing GroupNature Reviews Drug DiscoveryNektar TherapeuticsNeoGenix Oncology, Inc.Nerviano Medical SciencesNew England BioLabs, Inc.NextGen Sciences, Ltd.Nichirei BiosciencesNIH NCINIH NHGRINIH NIAIDNKT TherapeuticsNottingham City HospitalNovartis Horsham Research CenterNovartis Institutes for BioMedical ResearchNovartis Institutes for BioMedical Research, Inc.Novartis Pharma AGNovartis Vaccines & Diagnostics, Inc.Novexin, Ltd.Novo Nordisk ASNovo Nordisk ChinaNovo Nordisk Research USNovozymes Delta, Ltd.Nuvelo, Inc.Omrix BiopharmaceuticalsOncimmuneOncoMed PharmaceuticalsOrf GeneticsOrganogenesis, Inc.OrPro Therapeutics, Inc.Oswaldo Cruz FoundationPacifi c Biometrics, Inc.Pall Corp.Pall Life SciencesPanacea PharmaceuticalsParagon BioServices, Inc.Partners HealthcarePatrys, Ltd.PDL BioPharma, Inc.PerciviaPeregrine PharmaceuticalsPfi zer Global R&DPfi zer Global R&D Groton Labs

Pfi zer, Inc.Pharmaceuticals & Medical Devices AgencyPharmadule, Inc.PharmaNet Development GroupPhilip Morris InternationalPioneer Hi Bred International, Inc.Plexera BiosciencePotentia Pharmaceuticals, Inc.Premas Biotech Pvt, Ltd.ProGenosisPromega CorpProspect Venture PartnersProteos, Inc.Proteros Biostructures GmbHProtheonProtox TherapeuticsPublic Health Agcy of CanadaQIAGENQIAGEN GmbHQuanta BiosciencesRayBiotech, Inc.Regeneron Pharmaceuticals, Inc.Rigel Pharmaceuticals, Inc.Roche Discovery TechnologiesRoche Molecular BiochemicalsRoche Palo AltoRoyal Institute of Technology KTHsanofi aventis Grpsanofi pasteurSapidyne Instruments, Inc.Scarab Genomics LLCSchering PloughSCIL ProteinsScottish BiomedicalScripps Research InstituteSelexisSeraCare Life Sciences, Inc.SGX Pharmaceuticals, Inc.Shionogi Research LabShriners Hospital for ChildrenSidecSierra Sensors GmbHSloning BioTechnology, Ltd.SRU BiosystemsSt Jude Childrens Research HospitalStrategic BiosolutionsSyngentaSyntaxin, Ltd.Taisho Pharmaceutical Co., Ltd.TalecrisTalecris Biotherapeutics, Inc.Taligen TherapeuticsTandem LabsTeva Pharmaceutical Industries LTDTeva PharmaceuticalsThird Rock VenturesTibotec BVBATissue Transformation TechnologyTransgene SATransition Therapeutics, Inc.UCB CelltechUnited Biomedical, Inc.US Army Medical Research InstituteUS Environmental Protection AgencyVaccinex, Inc.VaxInnateVentana Medical SystemsVLST Corp.VybionWadsworth Center for Labs & ResearchWarwick Effect Polymers, Ltd.WyethWyeth Research LabsWyeth VaccinesXcellerex LLCXencor, Inc.XOMA US LLCZymeworks, Inc.ZymoGenetics, Inc.

Over 800 leaders gathered at the Fourth Annual PEGS meeting in Boston, Massachusetts from April 28-May 2, 2008. PEGS is the essential protein engineering summit and an event you can’t afford to miss.

Participating Companies in 2008 included:

PEGSummit.com 23

1. Registration Information ❒ Mr. ❒ Ms. ❒ Mrs. ❒ Dr. ❒ Prof. Name Job Title Div./Dept.CompanyAddressCity/State/Postal CodeCountryTelephone FaxEmail**Email is not a mandatory fi eld. However, by excluding your email you will not receive notifi cation about online access to pre-conference presenter materials, conference updates and networking opportunities.

Delivery Preferences: How would you prefer to receive notices from CHI: EMAIL: ❒ Yes ❒ No FAX: ❒ Yes ❒ No2. Pricing Information: Short Course Pricing

❒ SC1 Protecting IP for Protein Therapeutics and Diagnostics (Sun. 2-5pm)

❒ SC2 Phage and Yeast Display Libraries and their Screening (Sun. 10am-1pm) ❒ 1 Short Course ❒ $595 ❒ $295

❒ SC3 Small versus Large Molecule Therapeutics: Contrasting the Distinct Needs & Requirements for Biologics (Sun. 2-5pm) ❒ 2 Short Courses ❒ $895 ❒ $495

❒ SC4 Translational Strategies for Development of Monoclonal Antibodies from Discovery to the Clinic (Sun. 2-6pm) ❒ 3 Short Courses ❒ $1195 ❒ $695

❒ SC5 Dinner, Presentation and Interactive Panel Discussion: Focus on Aggregation: Mechanisms, Analytical Methods, and Real-World Examples (Thurs. 5:30-8:30pm)

❒ SC6 Testing for Immunogenicity (Sun. 2-5pm)

❒ SC7 Next Generation Sequencing Technologies for Antibody Clone Screening (Sun. 2-5pm)

Please select the package below based on the options you will most likely attend.

ADVANCE RATE until March 6, 2009 REGULAR RATE after March 6, 2009

❒ PREMIUM:(Includes access to conference options I, II, III)

❒ $2,550 ❒ $2,695

❒ $1,295 ❒ $1,445

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❒ $2,145 ❒ $2,345

❒ $1,075 ❒ $1,175

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3. Program Selections:Based on your pricing package, please select the programs you will most likely attend. NOTE: Choose one program per option.

CONFERENCE OPTIONS

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❒ Diffi cult to Express Proteins ❒ Immunogenicity of Therapeutic Biologics

❒ Scale-Up and Manufacturing

❒ Pre-Clinical/Clinical Development of Therapeutic Antibodies

❒ Proteins for Delivery ❒ Aggregation in Protein Therapeutics

Poster Discount ❒ $50 off

❒ I cannot attend but would like to purchase the PEGS event CD for $750 (plus shipping). Massachusetts deliveries will include 5% sales tax.❒ Please send information on exhibiting and sponsorship opportunities.

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Register by March 6th and SAVE! · the westin copley place boston the essential protein engineering summit PEGSFifth Annual final agenda April 6-10, 2009 PPhage Display of Antibodies