UH: QC

Quality Control in Regulated BiotechnologyUniversity of Houston1Quality ControlRaw Material, In-process, Final Product, Lot Release, and Stability

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Outline2DefinitionsRaw Material TestsIn-Process TestsFinal Product and Release TestsStability TestsSpecific Assays (commonly used assays)

Reference Q6B: Specifications : Test Procedures and Acceptance Criteria for Biotechnological/Biological Products @ www.ich.org

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Definitions3Contaminants materials unintentionally introduced during the process and not suppose to be in the final drug substance/drug productEx. Microbes, viruses, leachablesDrug product the protein in its final formulation in the labeled syringe or vial or other final packagingDrug substance the protein after the last purification step before fillingImpurities naturally occurring from the processEx. Cell culture media, host cell protein, host cell DNA, product related proteins (aggregates, misassembled protein, proteolytic fragments)Purity quantitative measurement of the desired protein

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Raw Materials Testing4Raw materials are biological or chemical substances used to manufacture the biopharmaceutical such as cell culture nutrients, serum components, inorganic salts, detergents, anti foam agents, enzymes, reagents, organic compounds, organic solvents, cleaning agents, growth factors, and chromatographic mediaThe basic quality concepts include assurance of the identity, purity, suitability and traceability of all the raw materials used in the manufacturing process. The quality of the raw materials used should meet standards appropriate for their intended use

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RM (contd)5Quality requirements for raw materials are met somewhat differently in production of clinical batches (process evolution) and manufacturing of licensed productThe requirements become stricter as you move from Phase I through Phase III and into commercial productionSpecial attention should be paid to raw materials of animal or human originThey should, as a general rule, be avoidedEach raw material should be evaluated to determine its criticality to the processRaw materials must be quarantined, identified, and released by an authorized person and their identity proven by specific assays (often available from the vendor)Certificates of analysis (COA) should be received for each lot of raw materialEach vendor must undergo a vendor qualification programIt is recommended, where possible, to use raw materials of USP or EP grade

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In-Process Testing6The safety and thereby the quality of biopharmaceutical drug products is linked to the process design, process robustness, process compliance with cGMP and extensive quality control programs including product specifications, process specifications, in process control, drug substance and drug product testing, regulatory policies and scientific understandingFor this class, discussion of in-process testing is limited to the analytical testing of intermediary compounds That is, samples taken anywhere during any process involved in making the final biological product

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7The purpose of the in process analytical control program is to ensure the quality of intermediary compoundsDuring initial development this part of the program is the most important part of process development as parameter intervals have not been defined at this early stageIt is common practice at this stage to make use of an extended analytical program, which is then later reduced as process validation progresses. Typical in process control analyses are test for microbial agents, test for fungi, test for mycoplasmas, test for viruses, test for endotoxins, 1D-SDS, HP-IEC, HP-RPC, HP-SEC, and ELISA

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Release Tests - Specifications8IdentityQuantity (related to potency)Physicochemical (related to identity)Purity, impurities and contaminantsPotency - biological activity (may include immunochemical properties)

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Identity9Assays which prove the protein is the correct protein and not a related one or another product made at the facilityThe assay should be unique for that protein.The test lot is run in the assay along with reference standardAcceptable assays - Peptide map, Ion exchange chromatography, ImmunoblotsInappropriate assays SEC, SDS gels

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Quantity10This is the measurement of the amount of drug substance/product.Normally A280 is used for proteins one of the most accurate measurements with S.D. 15% CV for multiple anlyte assays (host cell proteins)

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Immunoblots51Run the SDS gel colored molecular weight markersDo not fix or stain the gelElectrophoretically transfer the proteins from the gel to a PVDF membrane Wash and block the membraneAdd labeled antibody and washChemical reagent to visualize on film Line the membrane up with the film and mark where the colored MW proteins are

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Immunoblots - Westerns52

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Example of an Immunoblot53

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Q.C. Lab The Laboratory54Q.C. analysts trained on cGMP, general Q.C. SOPs, the instruments and the specifice SOP of an assayInstruments are validated IQ, OQ and PQCalibration and routine maintenance up to dateAssays may be new from development, qualified or validated

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