Transcript Document Systems Validation - MCCC Faculty & Staff Web Pages

QA Production Material
and Analytical Methods
BIT 230
Chapters 5 and 6 (Huxsoll)
Material Selection
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Original qualification of material vendor, specifications, safety, function,
etc.
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Lot-to-lot testing for release for
production use
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This chapter about original qualification
QA Materials
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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
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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
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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
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Safety first!
– Toxicity of extractables
– Qualification request:
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material to be approved
vendor identification
use
process conditions
general information
Personnel involved
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Requestor
Chemist
Biologist
Toxicologist
Safety Qualifications Manager
If pass test, the material can be
released to production
Suppliers
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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
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Meet timing needs (both material and
information requests about a material)
History with supplier
Technical knowledge
– had product for year or more
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Follow GMP guidelines
Size of supplier company
Good documentation with material
Type of Material Uses
 Nonproduct
 Product
contact
contact
Nonproduct contact
materials
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Engineering chemicals
– lubricants on equipment
– coolants (freon)
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Sterilants
– steam - most common one used - use WFI
for the steam
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Pesticides - not used in GMP facilityshould not have a pest problem!
Nonproduct contact
materials cont’d
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Paints
– solvents they emit- need ventilation (no
longer lead or mercury paints)
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Packaging
– not a big problem with nonproduct contact
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Colorants
– low toxicity colors such as white and ironoxide red
Product Contact Materials
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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
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Basic chemicals
– cell culture media
– water - most important raw material
– glass
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Process materials
– containers - glass or plastic (plastic needed
FDA approval)
Process materials cont’d
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Closures
Hoses and piping
– recommended: hard-piped stainless steel
– other types also acceptable (silicone)
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Affinity antibodies
– non-intentional components of productsmay leach from column, even though
should be tightly bound
– frequent washing of columns necessary
Process materials cont’d
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Pumps
– peristaltic pumps do not contain
extractables so ideal to use
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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
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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
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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
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Toxicity - in vitro biological reactivity test
– put material in fermentor with cells, look for
the material to damage or kill the cells
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Biological purity
– pyrogens
– viruses
– mycoplasma
– bacteria
Testing of process materials
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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
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Extracts testing
– Distilled water (DW) extractant for testing
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pH changes
oxidizable substances
heavy metals
nonvolatile residue
UV scan the DW extract
In process containers
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Stainless steel- has no toxic
components
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Glass - Type 1 accepted standard
– careful with high levels of aluminum in
glass
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Plastics- good properties - especially
polypropylene (what we use here)
Final containers
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Glass- same issues as with in-process
containers- need to be aware of
aluminum levels
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Plastics- low levels of extractables,
toxicity and aluminum, esp. PP (as long
as not repeatedly autoclaveddisposable)
Documentation
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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
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Microbiology - also initial and lot-to-lot
tests
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Toxicology - same as micro for tox tests
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Project Engineering - how exactly a
material is used in a process
QA of analytical methods
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Chapter 6
Biotech products
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Produced by either fermentation or cell
culture
Uses genetically engineered bacteria or
eukaryotic cells
Monoclonal antibodies - from hybridoma
cells
Proteins
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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
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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
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Therapeutic drug products
In vivo diagnostic testing
In vitro diagnostic testing
Medical devices
Agricultural products
Products for animals
Sterile injectable drugs
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This chapter’s focus
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Same 4 parameters - identity, quality,
purity and potency
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More tests on this type than othersmake sure they don’t pose a health risk
Physical tests
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Gross appearance
– color
– appearance
– pH
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Make sure there is no particulate matter
formed and precipitated
– provides stability information
Identity tests
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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
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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
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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
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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
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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
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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
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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
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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
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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
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GC
NMR
HPLC
Immunoassay
Remove chemicals near or below
detection limits
Other needs for purity
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Residual DNA
– reduce host cell DNA
– use hybridization assays
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Endotoxins
– use LAL test - see accompanying
presentation
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Mycoplasma
– broth and agar cultures
Potency tests
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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
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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
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Correlated results to WHO, NIH or USP
standards
measure in IU (international units)
Need consistent reagents
Well characterized reference standard
materials
Numerous replicates