ReaMetrix Overview

© 2008 Reametrix, Inc. All rights reserved.

ReaMetrixAn Introduction

2© 2008 Reametrix, Inc. All rights reserved.

Vision

ReaMetrix is committed to become a leading supplier of high quality and affordable integrated diagnostic solutions to resource poor settings, by leveraging local resources innovatively.


ReaMetrix at a glance

Life science technology company founded in May, 2003, CEO - Dr. Bala Manian, Founder of many life science companies

Strong management team with global experience in process and product development in regulated environments.

Started in San Carlos, CA with the centre of activity in Bangalore, India.

State of the art lab and production facility in Bangalore India.

Strong team, 42 employees, 38 in India, 15 with PhD’s.

ISO 9001:2000 Certified in May 2008 SAN CARLOS(HEAD

OFFICE)

BANGALORE(MANUFACTURI

NG & R&D

FACILITY)

SINGAPORE

(REPRESENTATIVE OFFICE)


Expertise in

• Gene design

• Cloning

• Expression

• Amplification & sequencing

Expertise in

• Tailoring assays to hardware platform

• Cell based and Bead based assays

• ELISA, FLISA

• From Proof of Concept to supply of assay kits

Expertise in

• Labeling of proteins and nucleic acids with

• Organic dyes

• Phycobiliproteins

• Nanocrystals

Expertise in

• Optics based instrumentation and detection systems.

• Integrated solutions with reagents to solve life science problems

AssayDevelopment

Stable CellLines

CustomConjugations Instrumentation

ReaMetrix Competencies

Core competencies drive Diagnostics & Research businesses


State of the art Indian Facility


PE TRCy5Cy5.5Cy7

Driving Value Through InnovationThe ReaMetrix Experience


Immunofluorescent Conjugates

Identifying bright & affordable fluorophores

Optimizing process chemistry to generate high performance conjugates

Innovation in design and process


Identifying Alternate Dyes – Developing a Palette

Phycoerythrin and the range of dyes in the red region PE emits at 562 nm Texas Red or equivalents emit at 610 nm Cy5,Dy649, Hilyte647 emit at 670 nm Cy5.5, Hilyte680, Dy680 emit at 700 nm Cy7 or Hilyte750, Dy750 emit at 770 nm

PE TRCy5Cy5.5Cy7

Allophycocyanin and the range of dyes in the red region APC emits at 660 nm Cy5.5 or equivalents emit at 700 nm Alexa700 or LiCor700 emit at 720 nm Cy7 or Hilyte750, Dy750 emit at 770 nm Licor800 or equivalents emit at 800 nm

APC Cy5.5A700 Cy7 Licor800

High Quality Fluorescent Dyes at Affordable Cost


Developing Immunofluorescent Conjugates

Optimize yields for fluorescent conjugations using process chemistry

Incoming Quality Control

Acquire unlabeled antibodies & other reagents from third

parties

Componentpurification

Functionalvalidation

Unitizing and packaging

Deliver finished product to customer

Specification/Quality AssurancePurification

Processquality control

testing

Conjugation

Process is our Product


Key features of ReaMetrix conjugates

Site Specific Labeling of Antibodies with precise Dye to Protein ratiosHigh-quality (certificate of analysis provided for each product with all the requisite specifications)

Validated for fluorescence intensity, activity, protein & dye concentration, dye integrity, emission spectra Perform functional validation where required

Audit trail (each and every step is documented to enable a high degree of transparency)Selected Customers who have validated ReaMetrix Conjugates

Harvard University, Duke University, National Institute of the Health, USA, Applied BiosystemsTata Memorial Hospital


Development conjugates for Leukemia-Lymphoma

Developed with 3 colors so that it can be used in India where 3 to 4 color Flow cytometers are prevalent.Conjugates have been validated on peripheral blood/bone-marrow samples or stable cell lines with receptors.The performance of these conjugates has been benchmarked with existing commercially available conjugatesReaMetrix Conjugates can be provided at smaller unit sizes so that they are affordable and accessible to more testing centers

S. No Antibodies FITC or eq PE PE-Cy5 eq APC PerCP1 CD2 Available Available2 CD3 Available Available Available3 CD4 Available Available Available4 CD5 Available Available5 CD7 Available Available6 CD8 Available Available7 CD10 Available Available8 CD11b Available Available9 CD11c10 CD13 Available Available11 CD14 Available Available12 CD15 Available Available13 CD16 Available Available14 CD19 Available Available Available15 CD20 Available Available Available16 CD22 Available Available17 CD23 Available18 CD25 Available Available19 CD33 Available Available20 CD34 Available Available21 CD36 Available22 CD38 Available Available23 CD41 Available24 CD45 Available Available Available Available25 CD56 Available Available26 CD5727 CD61 Available Available28 CD64 Available Available29 CD79a Available30 CD103 Available Available31 CD117 Available Available32 CD13833 HLA-DR Available Available34 Kappa Available Available35 Lambda Available Available36 Anti MPO Available37 Anti TdT38 FMC-739 IgM Available Available40 IgD41 TCR alpha/beta Available Available42 TCR gamma/delta Available43 Cyclin D44 Mouse IgG1-k (Isotype Control) Available Available Available45 Mouse IgG2a (Isotype Control) Available Available46 Mouse IgG2b (Isotype Control) Available

Coming Soon

Coming Soon

Coming Soon

Coming Soon

Coming Soon

Coming Soon

Coming Soon


Products Developed at ReaMetrix

Reagents for enumerating CD3+CD4+ reagentsTriTStat CD3/CD4/CD8TriTStat CD3/CD4/CD45ReaPan 34845ReaTcount

ReaPan B27 – Reagent for typing HLA B27ReaPan S34 – Reagent for enumerating CD34+ Hematpoietic stem cells

Value addition through Innovation


Typical Characteristics of ReaMetrix Products

Antibodies used are workshop approved clonesFluorescent labels used have no intellectual property entanglements.The assay protocols associated with these tests are similar to commercially existing assaysThe major difference is that ReaMetrix reagents are delivered in a unitized dry format.Particularly in resource poor settings, the dry reagent format reduces the cost of performing the assay and opens up additional options for providing access to the test for the patients.


Reagent for monitoring HIV progression through the Enumeration of CD3+CD4+ Cells

TriTStat CD3/4/8 and CD3/4/45MAH CD4 antibody labeled w/ Atto 488 dye (FITC alternative)MAH CD8 antibody (or MAH CD45) antibody labeled with R-Phycoerythrin (PE)MAH CD3 antibody labeled w/ PE-Dyomics 649 (Cy5 alternative) tandem dyePrecise number of fluorescent beadsStabilizers

Tests are suitable to run on open flow cytometers such as BD FACScan, BD FACSCalibur, Coulter EPICS etc.Reagents validated at

YRG care, ChennaiNARI, Pune

TriTStat CD3/4/8 is CE markedNow available as a Four color reagent as ReaPan 34845

Reagent supplied in Dry, Unitized Format No cold chain Up to 5 day stability of stained-fixed samples

CD4

0

300

600

900

1200

1500

0 300 600 900 1200 1500

Rmx Dry T-STATC

ou

lte

r T

etr

aC

hro

me r = 0.994


ReaTcount – Affordable Alternative to FACScount™

Reagent Supplied in a Dry, Unitized FormatReagent validated at

National HIV Immunology Laboratory, CanadaZimbabwe

CD4

0 100 200 300 400 500 600 7000

100

200

300

400

500

600

700

BD FACSCount

Rea

TC

ou

nt

Excellent Correlation


ReaPan B27 – For typing HLA B27

Use 2 clones – enhances sensitivity and specificitySelection of CD3+ population to reduce false positives Choice of PE as the dye to identify the HLA B27 population – High fluorescenceAddition of unconjugated Anti B7 antibody reduces nonspecific bindingProvision of Fluorescence Intensity beads in each reagent tube provides a built-in quality/reference standard

Dry Unitized Format Simplified work flow


ReaPan S34 – For Enumerating CD34 cells

Convenience: Dry, unitized ready to test format.Single platform enumeration using Modified ISHAGE guidelines.Stability: Dried down reagent does not require Cold Chain Maintenance.Workflow: Simple accurate and repeatable Lyse no wash protocol.Differentiates viable CD34+ stem cells from Non Viable(7-AAD).Single step pipetting : Minimizes errors.


New Product Ideas

Under development in our labs


ReaPanMicro – Presence/Absence Test for Bacteria in Urine

A diagnostic test to confirm the absence of bacteria in urine, based on the principle of nucleic acid detection in bacteria.Key Features of the Test

High sensitivity (100% Negative Predictive Value)Time to Result: Within 15 min of sample reaching a flow cytometer Easy to use, simple assay workflow

Culture Negative

Culture Positive146 Samples

By Culture

Overall TP TN

Positive by Flow 57 22

Negative by Flow 0 67

PPV 72.2%

NPV 100.0%


Bacteria Speciation Using Fluorescent Tags

Using Bacteriophages to detect bacteriaA: Bright field image of S.aureus bacteria stained with tag proteinB: Fluorescence image of stained S. aureus bacteria stained with tag proteinC: Bright field image of E. coli bacteria that was incubated with tag proteinD: Fluorescence image of E. coli bacteria that was incubated with tag protein

A B C D


Products under development - ControlsPositive Control Wells for Malaria

In collaboration with Foundation for Innovation in Diagnostics (FIND).To develop as a positive controls that will validate the efficacy of Rapid Diagnostic tests for Malaria

Blood ControlsTo develop blood controls for use in flow cytometry

http://www.finddiagnostics.org/news/newsletters/issue_10-jun_08/page03.shtml

Dot plots of 20 day stabilized blood samples


Products under development

ReaPan Leuco – To enumerate Leukocytes in Leukoreduced Platelets

For rare event detection and enumeration of leukocytes

HbA1CAn affordable method for %HbA1C using immunofluorescence

Viral vs Bacterial Pyrexia Differentiating between Viral and Bacterial Fever

LeukocytesBeads


Products under development

ReaPan CD61/41 – For the enumeration of platelets using immunofluorescent staining

For use in cases of Thrombocytopenia where existing enumeration methods do not work

ReaPan Mono – To enumerate Monocytes in peripheral blood.


ReaDev

Extending Innovations to Partners


Stable Cell Line Development

Stable CHOKI cells expressing IL8RBstained with fluorescently labeled chemokine IL8

Example of GPCR expressing cell lines developed at ReaMetrix


Bead Based Assay Development for US biotechnology Company

Develop Sandwich Bead Based assay for three different antigens with lower detection limits of 0.5 ng/mL

Transfer assay protocol with prototype kits (100 to 1000 tests/assay)

Working assay format with antibody coated carboxy beads and detection through Biotinylated Ab.SAV-PE successfully developed at desired detection limitsDelivered complete Antigen Detection Assay kits with detailed report including assay protocols

Antibody ScreeningAntibody Screening

Screened various commercially available antibodies (Ab) against analyte for best Kd

Screened various commercially available antibodies (Ab) against analyte for best Kd

Screened Antibodies in pairs to identify the pair with best detection sensitivity

Screened Antibodies in pairs to identify the pair with best detection sensitivity

Developed a bead based assay for the chosen analyte

Capture Ab on carboxy microsphere bead

Biotinylate detection Ab. Tertiary Detection Through SAV-PE

Developed a bead based assay for the chosen analyte

Capture Ab on carboxy microsphere bead

Biotinylate detection Ab. Tertiary Detection Through SAV-PE

Bead Based Assay Format

AntibodyPair Selection

AntibodyPair Selection Assay DevelopmentAssay DevelopmentR

EAMETRIX PROCESS

REAMETRIX PROCESS

Project Deliverables


Expression of cardiovascular protein for MAb development for US

Healthcare Company

Develop protocol for protein expression in mammalian cells.The protein is to be used for MAb development

Delivered purified protein at 1 mg/L expression level on optimized cell lineProtein expression and purification in native conformation without tags

Expression

Site directed mutagenesis Cloning into SK+ vector Sub-cloning into expression vectors Sequencing to confirm the clone

Site directed mutagenesis Cloning into SK+ vector Sub-cloning into expression vectors Sequencing to confirm the clone

Transfection and expression in three mammalian cell lines: CHO K1, COS 7, HEK 293

Transfection and expression in three mammalian cell lines: CHO K1, COS 7, HEK 293

Affinity chromatography Anion exchange chromatography Gel Filtration

Characterization SDS Page for Purity, Western Blot Analysis for Specificity, ELISA for quantitation

Affinity chromatography Anion exchange chromatography Gel Filtration

Characterization SDS Page for Purity, Western Blot Analysis for Specificity, ELISA for quantitation

Transfection Protein Purification & Characterization

REAMETRIX PROCESS

REAMETRIX PROCESS

Project Deliverables

Optimized Promoter/cell combinationDNA quantity per cellLipid to DNA ratioTransfection time

Lipids

DNA Lipid

complex

Cloned DNA

Secreted Protein


Towards Affordable Diagnostics

ReaMetrix Approach


Clinical Information Generation – Current Scenario

Sample journey and cost accumulation

Acquire Process Run Provide Results

Transport

Collection centre

Path LabDoctor

Hospital

Centralized Path Lab

Syringe/Vacutainer/Overheads

Cold chain/Courier

Tech Labor + Reagent

Capital Equipment

Amortization

Lab Infrastructure

Overheads

CD4 test example: ~10 to 20 times cost addition from reagent manufacture to cost to

patient Average cycle time for results ~ 48 to 72 hours

Layers of margin at every step


Issues with the current scenarioTechnical

Aged blood samples; accuracy & reliability of resultsTime lag for treatment decisionsLack of a patient centric approachImported equipment not designed for Indian environment. Eg. Automation, sample aggregation….

CommercialEncourages too many layers of margin affecting affordabilityCapital costs for imported equipment act as big barrier to entry as well as in adoption of closer to customer models

SocialAbsence of Economic value-add at the local level

Local economic activity needs to be promoted for enhanced reach of diagnostics


What is the ReaMetrix Solution?

Total systems approach – in developmentInstrument designed for local context robust & affordable.Reagents developed to suit local conditions temperature & humidity.Process improvement leveraging local talent Reducing the cost of the consumable kits.

Alternate business/distribution models enabling local empowerment.

From a clinical standpoint:Affordability enables more frequent testing & facilitates a more patient-centric approach.Accessibility facilitates rapid transmittal of information at the local level.

The ReaScan System – A Total Systems Offering


ReaScan – In Development


Summary

ReaMetrix is a platform for implementing innovative solutions to address the diagnostic needs of India

ReaMetrix is keen to work with partners who share a similar philosophy to create new solution opportunities


Manufactured by ReaMetrix India Pvt. Ltd.50-B, II Phase, Peenya Industrial Area

Peenya, Bangalore 560058, IndiaPh: +91-80-28378693/5, Fax: +91-80-

41172451E-mail: [email protected]

www.reametrix.com

Further details:

Health & Medicine

ReaMetrix Overview