Cell Processing: Validation & Performance

Qualification

Diane Kadidlo, MT (ASCP), SBBDavid McKenna, M.D.

PACT WebCastMarch 31, 2005

Cell Processing: Validation & Performance Qualification

Process ValidationWhat? Why? Who?

Validation vs. PQPerformance QualificationExample – Allogeneic NK Cells

From Discovery to Final Product

ValidationClinical

ProductionProcess

EvaluationBasic

ResearchTechnologyTransfer

ProcessDevelopment

• Clinical Indication

• Development Strategy

• Media Selection

• Bioengineering

• Characterization

• Media Optimization

• System Design

• Purification

• Practice Runs

• Process Development

• Equip/Supplies Qualification

•Documentation

• Clearance Studies

•Process Validation

• Contract Manufacturing

• Process Monitoring

• Stability Studies

• Outcome Review

Validation TypesEquipmentFacilityMaterialsPersonnelProductsComputerSystemTrainingLabelingPackagingSupplier & VendorProcess

Per FDA…

*Process validation is establishing documented

evidence which provides a high degree of assurance

that a specific process consistently produce a product

meeting it’s predetermined specifications and quality

attributes.

Process Validation What is it?

* Guidelines on General Principles of Process Validation May, 1987. FDA

Process Validation

Seunghwa Hong, Ph.D., Korea Food & Drug Administration, Presentation July 16, 2003.

Process ValidationWhy is it necessary?

Ensure compliance with regulations (GxPs) and internal policies and procedures

Ensure effectiveness, purity, quality and safety of product produced

Ensures that the item validated functions accurately and consistently as intended

Process ValidationWho is responsible?

Executive Management: Upper Management, Directors, CEO

Quality Assurance Unit: Generally has oversight of the

entire validation program for a facility

Process Owners: The staff performing the validation

Government Agents: The FDA has the role of ensuring

validations are occurring

Validation vs. Qualification

ValidationLife cycle of a product (i.e., development use

maintenance)Cell processing laboratories are responsible for validating

own processes (personnel, equipment, methods, SOPs) to ensure compliance with cGMPs/cGTPs.

QualificationPart of validation process which verifies functional

performance prior to clinical production and thereafter according to SOPs.

Process Validation

Seunghwa Hong, Ph.D., Korea Food & Drug Administration, Presentation July 16, 2003.

Performance Qualification Prerequisites

Before writing cell processing PQ protocol:

Support: QA and financial supportPlan: schedule, budget Equipment: calibration & maintenanceSupplies & Materials: vendor qualification, C of AsPractice runs

Cell Processing PQ Protocol

Authorized document that defines your plan for PQ including:Product typePurpose & rationaleProcess (reference to Batch Record/SOP/Worksheets)Reagents/Supplies/EquipmentTimetableNumber of runs (general rule of minimum of 3)Acceptance criteriaMethod of analysisRevalidation criteria (run failure, changes)Approval: Lab Director, QA, others: PI, Manager

Acceptance CriteriaSafety

Donor infectious disease testingGram stainSterility cultures (USP/clinical laboratory)Mycoplasma detection (indirect/direct)Testing for adventitious agents

PurityEndotoxinAssays for assessment of in-process reagent removal (e.g., antibodies/ beads, vectors, cytokines)

IdentityFlow cytometry analysis CytochemistryMolecular diagnostic testing (e.g., quant. RT-PCR)

PotencyViability (microscopy/flow cytometry)Clonogenic Assays (LTC-IC, CFU, SRC)Proliferative potential (e.g., G-banding, Southern blot for telomere length)Pluripotency (in-vivo differentiation potential in NOD-SCID mice)

StabilityAs noted for ‘Potency’ after short- and long-term storage

Performance Qualification

Execute Validation

Review data

Review Deviations/ChangesReviewEvaluate whether event impacts validation outcomeRevalidate if appropriateDocument

Summary Report

Performance Qualification

Allogeneic Natural Killer Cells

QOD IL-21.75 MU/m2 s.c.

-7 -6 -5 -4 -3 -2 -1 0 14

2 x 10 7/kg

Cyclophosphamide60 mg/kgAML only

Fludarabine25 mg/m2

Allogeneic NK cells

• Collectdonor cells

• T-cell deplete

• Activatewith IL-2

In vivo expansionRes

t

Clinical Trial

76.4 %76.4 %TT--cellscells

0.1 %0.1 %TT--cellscells

PBMC Apheresis

CD3+ cell depletion

Activation/Expansion

Infuse

Harvest

IL-2/X-VIVO-15

Suspend in NS/5% HSA

37% NK cells

Conclusions

Allo-NK cells expand in vivo; evidence of efficacyLarger cohorts needed to fully demonstrate efficacyMore pure NK cell product (no B cells and lesscompeting T-cells) may increase safety and efficacy

Cell Processing PQ ProtocolAllogeneic NK Cells

Product type: Allogeneic NK cellsPurpose & rationale: Provide more pure product (safety, efficacy)Process: Established; Additional purification stepNumber of runs: minimum of 3 consecutive, successful runs Acceptance criteriaMethod of analysisApproval: Lab Director, QA, others: PI, Manager

Acceptance Criteria

Assay Test Method Specification

Viability Flow Cytometry > 70%

NK Cell Enumeration Flow Cytometry >70%

T Cell Enumeration Flow Cytometry < 3 x 104/kg

Endotoxin LAL Method < 5 EU/kg

Gram Stain Routine No organisms

PBMC Apheresis

CD3+ cell depletionCD56+ cell enrichment

Activation/Expansion

Infuse

Harvest

IL-2/X-VIVO-15

Suspend in NS/5% HSA 92% NK cells

• Results from PQ summary report (Table)

Summary

Process Validation: Assurance of process consistency; full life cyclePerformance Qualification: Part of validation; verification of functional performancePQ Protocol: Essential to define plan