Current Trends and Strategies in Analytical Method Validation, applied to Biologicals and Cell Therapy products R. Jans Mobile 0492 593602 Email [email protected]Sint-Lambertusstraat 141 Rue Saint- Lambert 1200 Brussels www.businessdecision-lifesciences.com
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QC Method Validation for Biologicals vs Cell Therapy products (ATMP)
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Current Trends and Strategies in
Analytical Method Validation, applied to Biologicals and Cell Therapy products
Methods required for QC release of biologicals & cell therapy products
• Microbiological Testing– straightforward
• Identity• Purity / Content• Potency• Stability
– uses ID/content and potency assays
Ref: FDA Guidance for Industry: Content and Format of Chemistry, Manufacturing and Controls Information and Establishment Description Information for a Vaccine or Related Product (1999)
Ref: FDA Guidance for Industry: Guidance for Human Somatic Cell Therapy and Gene Therapy (1998)
Ref: Guidance for FDA Reviewers and Sponsors: Content and Review of Chemistry, Manufacturing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs) (2008)Ref: FDA Guidance for Industry: Potency Tests for Cellular and Gene Therapy Products (2011)
Example: validation of ID/Content ELISA assay for vaccine X
• ELISA– specificity (matrix + antigen)– linearity-precision-accuracy-range:• 2 ops, n=2, 4 days-> n=16 par level• 8 levels of validation standard, 50%-150% of specification: diluted FC,
FC, FC+spikes with standard, spike only• data n=128• ANOVA
– robustness– stability-indicating
Differences in QC release testing methods for biologicals and cell therapy products
Test Type Biologicals Cell therapy product
Identity(What is it?)
Physico-chemical and biochemical methods: NMR, HPLC, ELISA,
Western blot, ...
Biomarkers by biological methods: flow cytometry, qPCR, ...
Purity(How pure is it?)
HPLC, ...
Non-cell impurities: Physico-chemical and biochemical
methodsCell population purity: Biomarkers by
biological methods
Potency(How efficacious
is it?)
In vivo or surrogate biological or biochemical methods (content)
In vivo or surrogate biological or biochemical methods
Challenges for identity/content method validation, biologicals vs.cell therapy products: WHAT IS IT?
• biologicals, for example protein vaccine– identity/content by ELISA– specific GMP antibodies– GMP conform reagents– GMP reference standard– straightforward validation
• cell therapy products, for example MSC-type product– identity by multicolor Flow Cytometry– limit-type assay– desired cell type = MSC primed for specific differentiation pathway– biomarkers? – antibodies/reagents? (GMP)– validation of a flow cytometry method?
The nemesis of validating QC release methods for cell therapy products
Challenges for purity testing, biologicals vs.cell therapy products
• biologicals, for example protein vaccine– impurity dosage by physico-
chemical testing– quantitative assays– GMP conform reagents– GMP reference standard– straightforward validation
• cell therapy products, for example MSC-type product– purity by qPCR– limit-type assay– undesired cell types = mesenchymal stem cells differentiating towards
bone, fat or cartilage– biomarkers? – qPCR primers?– validation of a qPCR method?
Challenges for potency testing, biologicals vs.cell therapy products
• biologicals, for example protein vaccine– potency testing in vivo
• inject vaccine in vivo anddose serological response using ELISA
• « quantitative « assay• variability = nightmare
– potency testing in vitro• ELISA• relatively straightforward validation
• cell therapy products, for example MSC-type– desired effect: tissue regeneration – potency testing in vivo
• technically not feasible for autologous batches
– potency testing in vitro• surrogate markers for biological activity - bioassays
– engraftment and differentiation into functional tissue– paracrine secretion of regenerative proteins
Conclusions
• QC method validation for biologicals & cell therapy products– evolving field – requiring scientific creativity– involves latest technology– constant exchange of ideas with regulatory