Manufacturing and Quality Control of Bacterial Products

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Lecture No. 15 - April 16th, 2004 Manufacturing
Quality Control of Veterinary Vaccines Robby
Robinson
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Manufacturing and Quality Control of Veterinary
Vaccines

• Dr. Robby Robinson
• Pfizer Global Manufacturing
• Lincoln Operations

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Veterinary Vaccine Production

• Product design
• Master reagents
• Growth harvest
• Preparation of the product
• Testing
• Post-preparatory steps
• Documentation
• Economics

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Product Design

• Value added Efficacious, safe,
  convenient
• Claims
  Mfg QC must satisfy
• Country
  Reg. systems add challenges
• Product type Live,
  killed, combination
• Bacterin, toxoid, vaccine, subunit
• Production systems Bact, viral, in vivo, in
  ovo, plant
• Presentation and packaging

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Master Reagents

• Starting points
  Cornerstones
• Loss Redevelopment Manage!
• Master seeds
• Master cell stocks
• Master QC reagents
• Challenge strains

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Master Reagent Concepts

• Proprietary
  Patented?
• Genetic purity is better Cloning
• Must protect
  Planning
• Will be retested challenged

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Master Seed Organisms

• Strain Heterogeneic?
  Contaminated?
• Source Country, animal, individual,
  institution
• History Cells, animals, location,
  materials
• Identification Biochemical, serological,
  antigenic
• 
  nucleic acid sequence

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Master Seed Organisms

• Genetic stability
• Balance between safety efficacy for attenuated
  organisms
• Attenuation
  Classical or rDNA
• Passages permitted Usually 5 - 7
• Stability during storage

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Master Cell Stocks

• Pure?
• Identity
  Karyology, FA
• Genetic stability Usually 20 passages
• Purity Bacteria,
  viruses, myco, cells
• Stability Liquid vs. vapor
  N2 storage

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Master QC Reagents

• Monoclonal antibodies Hybridomas
• References
  Requalification
• Antisera Identification
• Test strains?
• Controls?

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Raw Material Sourcing

• Pieces parts of dead animals
• Global marketplace
• Wide variability in quality
• Potential for disease spread
• BSE concerns
• Growth media components
  (serum, hydrolysates, vitamins, extracts)
• Stabilizers (gelatin,
  hydrolysates)
• Adjuvants (oils,
  surfactants)
• Must be managed

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Raw Material Challenges
Validated Effectiveness of the Process
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Growth and Harvest

• Logistics
  Unappreciated
• Materials
  Ingreds of animal origin
• Equipment Expensive,
  very technical
• Variability
  Ability to respond limited
• In-process specifications Sterile, potent
• In-process testing Significance?
• Documentation and records QA

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Preparation of the Product

• Inactivation Formalin, BPL, BEI,
  merthiolate
• Downstream processing Clarification, conc,
• 
  diafiltration, extraction
• Detoxification / Purification Endotoxin
• Standard testing Nephelo, potency, sterility,
  myco,
• 
  extraneous, inactivation
• Toxoiding
  Formalin, heat, pH
• Standardization Extenders, potency
  testing,
• 
  consistency (safety efficacy)

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GMP Inactivation Model
Kinetic Analysis Std Conditions
3X Inactivation Procedure
Inactivant Conc. Selection


Master Cells
Validated Agn Prod Method
Live Virus Harvests
Master Seed
Reagents
Detection System (eg, FA Cell)
Valid Inact Test
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Inactivation
Constant Conditions 22 1O C 48 2 hr.
Inactivant Concentration
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Preparation of the Product

• Adjuvanting AlOH, emulsions,
  saponins
• Mixing Time, pH, settling,
  resuspension
• Bulk testing Potency, pH
• Filling Equipment vial sizes,
  campaigns
• Lyophilization Stability at a cost,
  subserials
• Antigen/dose doses/container

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Testing

• Interwoven at key stages
• Purity Sterility, mycoplasma,
  extraneous virus
• Safety Host and/or lab animal
• Potency Not enough? Too much?
• Efficacy Will it work? Indirect
  measurement

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Extraneous Agent Testing Limitation

• Extraneous agent testing can only detect
• Heavily contaminated raw materials
• Bugs that have replicated in a production system
  after contamination at an earlier point
• A misassembled final product
• It cannot reliably detect low levels of
  contamination
• in a sample due to simple probability and
  technical difficulties associated with many
  detection systems.

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Virus Contamination Sources

• Extraneous Virus Sources
• Bio Raw Materials
• Faulty Equipment Decontamination
• False-Neg Test Results
• Master Seeds Cells
• Human Error
• Incomplete Inactivation

Its Seldom Cross- Contamination!
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Documentation (1)

• Regulatory requirements 9CFR, etc
• Outline of Production
• Standard Operating Procedures
• Records Mfg worksheets, QC test records
• Validation Equipment,
  processes, tests
• Raw materials Specs
  testing

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Documentation (2)

• Animal procurement holding
• Inventory records
• Sales records To customer
• Personnel training Sampling
• Final releases USDA, QA
• Documentation is a product

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Economics 101

• Development cost
• Sales price
• Cost of goods sold (CGS)
• 
  Materials, labor and overhead
• Operating Sales Administration
• Profitability (Sales - CGS - Operating) /
  Sales
• High cost No sales No job