qa production material and analytical methods bit 230 chapters 5 and 6 (huxsoll)
TRANSCRIPT
QA Production Materialand Analytical Methods
BIT 230
Chapters 5 and 6 (Huxsoll)
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
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
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
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
Qualification of Materials
Safety first!– Toxicity of extractables– Qualification request:
• material to be approved• vendor identification• use• process conditions• general information
Personnel involved
Requestor Chemist Biologist Toxicologist Safety Qualifications Manager
If pass test, the material can be released to production
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
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
Type of Material Uses
Nonproduct contact
Product contact
Nonproduct contactmaterials
Engineering chemicals– lubricants on equipment– coolants (freon)
Sterilants – steam - most common one used - use WFI
for the steam Pesticides - not used in GMP facility-
should not have a pest problem!
Nonproduct contactmaterials 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 iron-
oxide red
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
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)
Process materials cont’d
Closures Hoses and piping
– recommended: hard-piped stainless steel– other types also acceptable (silicone)
Affinity antibodies– non-intentional components of products- may
leach from column, even though should be tightly bound
– frequent washing of columns necessary
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
Testing of materials
Plastics
–material must pass USP testing
Closure
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
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)
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
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)
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
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)
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 autoclaved- disposable)
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
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
QA of analytical methods
Chapter 6
Biotech products
Produced by either fermentation or cell culture
Uses genetically engineered bacteria or eukaryotic cells
Monoclonal antibodies - from hybridoma cells
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)
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
Uses of biotech products
Therapeutic drug products In vivo diagnostic testing In vitro diagnostic testing Medical devices Agricultural products Products for animals
Sterile injectable drugs
This chapter’s focus
Same 4 parameters - identity, quality, purity and potency
More tests on this type than others- make sure they don’t pose a health risk
Physical tests
Gross appearance– color– appearance– pH
Make sure there is no particulate matter formed and precipitated– provides stability information
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 78-80- will go over
more in next lecture
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
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
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
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
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
Immunoassays
Determines identity (by reacting with specific antibody)
Determines specific activity (when uses as part of total protein measurements)– RIA– ELIZA– IRMA (immunoradiometric assays)
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 6.1 - test with
sensitivity ability
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
Added chemicals cont’d
GC NMR HPLC Immunoassay Remove chemicals near or below
detection limits
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
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
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
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