QA Production Material and Analytical Methods. BIT 230 Chapters 5 and 6 (Huxsoll)

Transcription

1 QA Production Material and Analytical Methods BIT 230 Chapters 5 and 6 (Huxsoll)

2 Material Selection Original qualification of material - vendor, specifications, safety, function, etc. Lot-to-lot testing for release for production use This chapter about original qualification

3 QA Materials Start in R & D - in large companies, sometimes R & D personnel don t know that QA group should be aware of their materials Need to know QA materials need testing and approval

4 Technology Technology groups (a.k.a. tech transfer)- transfer the process from research to development to production (manufacturing technology or process development Perform scale-up procedures Don t always use initial research materials in production

5 Engineering Keeps process going once a material is in use Need to know specs of required materials (equipment parts, e.g.) Approve new materials or parts before installation Work from approved vendors

6 Qualification of Materials Safety first! Toxicity of extractables Qualification request: material to be approved vendor identification use process conditions general information

7 Personnel involved Requestor Chemist Biologist Toxicologist Safety Qualifications Manager If pass test, the material can be released to production

8 Suppliers For start up biotech (and maybe large pharma too?) use biggest, most experienced, technically sound suppliers Don t want to find a guinea pig here Small co. can t do a lot of their own testing, so have to reply on tried and true vendor

9 Considerations when choosing a Supplier Meet timing needs (both material and information requests about a material) History with supplier Technical knowledge had product for year or more Follow GMP guidelines Size of supplier company Good documentation with material

10 Type of Material Uses Nonproduct contact Product contact

11 Nonproduct contact materials Engineering chemicals lubricants on equipment coolants (freon) Sterilants steam - most common one used - use WFI for the steam Pesticides - not used in GMP facilityshould not have a pest problem!

12 Nonproduct contact materials cont d Paints solvents they emit- need ventilation (no longer lead or mercury paints) Packaging not a big problem with nonproduct contact Colorants low toxicity colors such as white and ironoxide red

13 Product Contact Materials Two main types: Basic chemicals (put into process intentionally) Process materials (may be by-products of the process)- does not bind up product

14 Product Contact Materials cont d Basic chemicals cell culture media water - most important raw material glass Process materials containers - glass or plastic (plastic needed FDA approval)

15 Process materials cont d Closures Hoses and piping recommended: hard-piped stainless steel other types also acceptable (silicone) Affinity antibodies non-intentional components of productsmay leach from column, even though should be tightly bound frequent washing of columns necessary

16 Process materials cont d Pumps peristaltic pumps do not contain extractables so ideal to use Gaskets, O-rings etc. rubber problematic in biotech production problems with sticking need to verify integrity with toxicity testing

17 Testing of materials Plastics material must pass USP testing Closure

18 In-house testing Not just vendor specs, but need to establish in house specs for raw materials USP the basis Other tests - chemical, physical, Chemistry, toxicology and microbiology involved in determining tests

19 Parameters to check Basic chemicals appearance identity (IR spec) Assay (HPLC) Purity - need to define impurities first; also use HPLC, or TLC (thin layer chromatography)

20 Parameters to check cont d Toxicity - in vitro biological reactivity test put material in fermentor with cells, look for the material to damage or kill the cells Biological purity pyrogens viruses mycoplasma bacteria

21 Testing of process materials As is testing Identity - IR Reside on Ignition (ROI) - solid is ashed at 600 C; ensures supplier is using same inorganic raw materials Emission Spectrographic Analysis (ESA) - residue from ROI tested for heavy metals (cadmium or lead, for example)

22 Testing of process materials cont d Extracts testing Distilled water (DW) extractant for testing ph changes oxidizable substances heavy metals nonvolatile residue UV scan the DW extract

23 In process containers Stainless steel- has no toxic components Glass - Type 1 accepted standard careful with high levels of aluminum in glass Plastics- good properties - especially polypropylene (what we use here)

24 Final containers Glass- same issues as with in-process containers- need to be aware of aluminum levels Plastics- low levels of extractables, toxicity and aluminum, esp. PP (as long as not repeatedly autoclaveddisposable)

25 Documentation Departments involved in release of materials after validation: Purchasing - request vendor audits Inspection and Receiving - know about arrival of material and how to handle Safety and Environmental Chemistry - choose tests for initial evaluation and lot specific tests

26 Documentation cont d Microbiology - also initial and lot-to-lot tests Toxicology - same as micro for tox tests Project Engineering - how exactly a material is used in a process

27 QA of analytical methods Chapter 6

28 Biotech products Produced by either fermentation or cell culture Uses genetically engineered bacteria or eukaryotic cells Monoclonal antibodies - from hybridoma cells

29 Proteins Similar properties - therefore need to develop methods to characterize them High molecular weight 10,000-20,000 Daltons (insulin and interferon) > 950kD (IgM monoclonal antibodies)

30 Proteins This chapter will summarize- more about proteins in next lecture Further testing parameters for proteins: primary, secondary, tertiary & quaternary disulfide bonding multiple chains carbohydrate content

31 Uses of biotech products Therapeutic drug products In vivo diagnostic testing In vitro diagnostic testing Medical devices Agricultural products Products for animals

32 Sterile injectable drugs This chapter s focus Same 4 parameters - identity, quality, purity and potency More tests on this type than othersmake sure they don t pose a health risk

33 Physical tests Gross appearance color appearance ph Make sure there is no particulate matter formed and precipitated provides stability information

34 Identity tests Molecular weight retention times peptide mapping reaction with an antibody biological activity in assays N- and C- terminal sequence analysis Review each one pages will go over more in next lecture

35 Assays Quantify proteins Total protein content Amino acid composition Degradation products Measure of glycosylation (carbohydrate content) HELPS measure lot-to-lot consistency of a biotech product

36 Total protein assays Lowry, Bradford, etc- (one we did) Just measure total protein (not specific) Need external reference for each test Bradford uses Coomassie blue dye (one we used- the darker the blue color the greater amount of protein) Each assay requires spectrophotometry measurements

37 Native protein Protein can lose its native form easily - by fragmentation, aggregation, denaturation, or chemical modifications Use separation methods to remove from native proteins Use HPLC, SDS-PAGE

38 Amino acid comp. analysis Quantitate amount of each a.a. in protein Used especially to identify after a manufacturing change occurs Routine control test, too Digest protein into a.a. componments

39 Carbohydrate analysis No glycoproteins in bacteria; found in proteins produced in eukaryotic cells Sugars found include galactose, glucose and mannose Individual sugar content determined by HPLC or GC

40 Immunoassays Determines identity (by reacting with specific antibody) Determines specific activity (when uses as part of total protein measurements) RIA ELIZA IRMA (immunoradiometric assays)

41 Purity tests Affected by contaminants and degradation products that co-purify with protein during production WCB can be source of contaminants Purification accomplished by chromatography See page 84 Table test with sensitivity ability

42 Added chemicals Chemicals added during fermentation, cell culture and protein purification How do you then get rid of the chemicals from the final product? End product testing required - many methods can be used to test absence of added chemicals in final product

43 Added chemicals cont d GC NMR HPLC Immunoassay Remove chemicals near or below detection limits

44 Other needs for purity Residual DNA reduce host cell DNA use hybridization assays Endotoxins use LAL test - see accompanying presentation Mycoplasma broth and agar cultures

45 Potency tests Measure dose-dependent biological activity Designed to mimic in-vivo activity of the biological product Performed in animals or in cellular assays Cellular assays ideal over animal assays

46 Potency tests cont d What is measured: protection from viral infection clot dissolving properties binding to specific antigens inhibition of protein synthesis inhibition of DNA replication

47 Potency tests cont d Correlated results to WHO, NIH or USP standards measure in IU (international units) Need consistent reagents Well characterized reference standard materials Numerous replicates