QC Method Validation for Biologicals vs Cell Therapy products (ATMP)
Current Trends and Strategies in Analytical Method Validation, applied to Biologicals and Cell Therapy productsR. JansMobile 0492 593602Email firstname.lastname@example.orgSint-Lambertusstraat 141 Rue Saint-Lambert1200 Brusselswww.businessdecision-lifesciences.com
Analytical Method Validation for Beginners• Validation ? – GMP: Validation is the documented act of demonstrating that a procedure, process, and activity will consistently lead to the expected results.• Validation of analytical methods for QC? – Validation of an analytical method is the process by which it is established by laboratory studies, that the performance characteristics of the method meet the requirements for the intended analytical application• Main Message: Method Validation ... – ... for „classical‟ pharmaceutical compounds = „easy‟ & straightforward • chemistry-based – ... for biologicals = scientific creativity required, constant exchange of ideas with authorities • biochemistry- and biology-based – ... for cell therapy products = biologicals3
Analytical Method Validation for Beginners• For a qualitative assay: – test result = pass or fail, no quantitation – validate specificity and detection limit• The “10 commandments” for a quantitative assay: 1. specificity 2. linearity 3. accuracy 4. precision 5. range 6. detection limit 7. quantitation limit 8. robustness 9. stability-indicating 10.system suitability test
Biologicals & Cell therapy products• Biologicals – washing powder – typically pharmaceutical products • of biological origin – vaccines » live or fixed viruses or bacteria (rota, polio, hepatitis A, whole cell pertussis, ...) • bio-engineered – vaccines » virus-specific proteins produced by genetically engineered bacteria (HBsAg) » virus- or bacteria-specific polysaccharides produced by genetically engineered bacteria – humanized antibodies » anti-psoriatics, Enbrel or Remicade (anti-TNFa)
Cell therapy products• Cellular and Gene Therapy products (FDA)• Advanced Therapies Medicinal Products (EMA)• Pharmaceutical product = Cells – Haematopoietic stem cells for restoring bone marrow function after cancer treatment – minimally manipulated – Regenerative Medicine • Stem Cells, autologous • Stem Cells, allogeneic – Anti-cancer cell therapy • Allogeneic Lymphocytes • Allogeneic Dendritic cells
Methods required for QC release of biologicals & cell therapyproducts Ref: FDA Guidance for Industry:• Microbiological Testing Content and Format of Chemistry, Manufacturing and Controls Information and Establishment – straightforward Description Information for a Vaccine or Related Product (1999)• Identity Ref: FDA Guidance for Industry: Guidance for Human Somatic Cell• Purity / Content Therapy and Gene Therapy (1998) Ref: Guidance for FDA Reviewers• Potency and Sponsors: Content and Review of Chemistry, Manufacturing, and• Stability Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug – uses ID/content and potency assays Applications (INDs) (2008) Ref: FDA Guidance for Industry: Potency Tests for Cellular and Gene Therapy Products (2011)
Example: validation of ID/Content ELISA assay forvaccine 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 andcell therapy products Test Type Biologicals Cell therapy product Physico-chemical and biochemical Identity Biomarkers by biological methods: methods: NMR, HPLC, ELISA, (What is it?) flow cytometry, qPCR, ... Western blot, ... Non-cell impurities: Physico-chemical and biochemical Purity HPLC, ... methods(How pure is it?) Cell population purity: Biomarkers by biological methods Potency In vivo or surrogate biological or In vivo or surrogate biological or(How efficacious biochemical methods (content) biochemical methods is it?)
Challenges for identity/content method validation, biologicalsvs.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 therapyproducts
Challenges for purity testing, biologicals vs.cell therapyproducts• 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 therapyproducts• biologicals, for example protein vaccine – potency testing in vivo • inject vaccine in vivo and dose 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 authorities