1. What Limits the Rate of Vaccine Development and Production? Alan R. Shaw VaxInnate Corporation

2. Vaccine R&D in one slide If natural infection results in protection… –Mimic the natural response as best you can If natural infection is not resolved by the host immune response… –Prepare for a long and difficult program

3. Assumptions for today Target antigen(s) defined Method of presentation defined –Live attenuated virus –rDNA VLPs –Vectored gene delivery –Soluble proteins or peptides on arrays Biological manufacturing “host” defined –Mammalian cells, eggs, yeast, insect cells, prokaryotes, tomatoes, etc.

4. Now that we know what we’re making and how… Biggest job is analyzing what you’ve made Consistency of product among batches –Potency –Stability –Purity –What aspects of your product and process are relevant to these attributes? –How much variability is allowable (as opposed to measurable)?

5. Analyticals Each test must be validated –Sensitivity, limits of detection, quantitation –Reproducibility, inherent variability –Robustness, reagents, operators, days Equipment associated must be validated –Software, and hardware

6. Analyticals Support Process Development Circular, iterative process Analytical tools refined as process evolves A good, stable process depends on good, stable analytical tools Selected assays will be used for release of licensed product –Internal QC, FDA/CBER release

7. Similar Analytical Exercise for Clinical Trial support Identify most relevant aspect of immune response Hope that this can be measured in an accessible sample Validate performance of assays and equipment Also applies to detection of pathogen

8. Impact of Timing Assays need to be “ready” in order to proceed This can be a rate-limiting step!!! Especially troublesome when assay result does not tell you what you think it’s telling you –Does this track with stability? –Does this track with potency?

9. From Pilot Plant to Manufacturing Pilot facilities mimic process flow but not scale Typically, 10X jump from pilot to full-scale production Issues are often heat transfer, oxygen transfer, nutrient supplies, odd changes in host cell physiology at scale Having Process R&D and Manufacturing in one place is helpful

11. Highly Coordinated Effort Stage 4 Review Proof of Concept Stage 0 Product & Process Definition Stage 1A Dose & Scale Definition Stage 1B Proof of Efficacy & Manufacturability Stage 2 WMA Preparation Stage 3 License & Launch Stage 4 Marketing Clinical/Regulatory Process Analytical Assays Serology Assays Product Definition, Packaging & Stability Supply / Production Marketing Needs Report Pre-Launch Strategy Worldwide Marketing Plan Proof of Concept- Clinicals Product, Process and Formulation Development Process Support and Optimization Validate Potency, Safety & Ster. Assay Validate Raw Material, Stability and Release Assay Evaluate Assay Performance Develop & Evaluate High Throughput Serology Assay Manufacturing Feasibility Studies Economic Feasibility Assessment Mfg.. Strategy Prepare Prelim. Eng.. & Basis of Design Inspection Preparation Prepare Lab Lots for POC Clinicals Prepare Pilot Lots for Dose Ranging Prepare Phase 3 Quality Lots Prepare Consistency Lots Prepare Launch Quantities of Vaccine Start Stage 0Stage 0 ReviewStage 1A ReviewStage 1B ReviewStage 2 ReviewStage 3 Review Dose Defined Regulatory Assessment Proof of Concept Bridging Studies Dose Ranging Studies Process Assessment- Clinicals Efficacy Studies Consistency Studies Scale Defined LPO Submit IND Prepare WMA Support WMA & Extend Expiry First Sale Release WMAApproval Product, Process & Formulation Finalized Kit Vendor Development of Serology Assays Capable of Evaluation Post-Vaccination and Post-Disease Seroconversion ID Parameter for POC Clinicals Stage 1 Anal.& Process Plan Spec.. Strategy & Rationale Validate In-Process and Product Characterization Assays Update Spec. Strategy & Rationale Transfer Assays Update Spec. Strategy & Rationale Update Spec. Strategy & Rationale Detail Design Order Equipment Build and Validate Facility Statement of Interest (SOI) RMC Approval Facilities Early Research Stability Experiments Preliminary Product & Packaging Definition Probe & Clinical Lot Stability Studies Pilot Lot Stability Studies Final Product & Primary Package Definition Post-Launch Product and Packaging Support Filling and Packaging Development Launch and Annual Stability Studies Full Scale Lot Stability Studies Market Container Stability Studies Final Secondary Package Definition

12. Fermentation Suite

13. Biology is Also Rate-Limiting Biomass expansion is determined by doubling time of host cell Release testing is limited by physiology of the test system

14. Two Divergent Examples Varicella vaccine Attenuated virus Made in MRC5 cells 24 hour doubling time ~200 doses per roller bottle 48 hour infection cycle (x2) VaxInnate’s flu vaccine Soluble recombinant protein Made in E. coli 50 MM doses per 2000L tank 20-30 minute doubling time

15. Divergent Testing Varicella –Egg safety –Sterility –Potency (plaque) –Suckling mouse –Guinea pig –Tissue culture –Mycoplasma (x2) –Residual moisture rDNA influenza –Sterility –Potency, ELISA –Dose, HPLC –excipients

16. Highly Coordinated Effort Stage 4 Review Proof of Concept Stage 0 Product & Process Definition Stage 1A Dose & Scale Definition Stage 1B Proof of Efficacy & Manufacturability Stage 2 WMA Preparation Stage 3 License & Launch Stage 4 Marketing Clinical/Regulatory Process Analytical Assays Serology Assays Product Definition, Packaging & Stability Supply / Production Marketing Needs Report Pre-Launch Strategy Worldwide Marketing Plan Proof of Concept- Clinicals Product, Process and Formulation Development Process Support and Optimization Validate Potency, Safety & Ster. Assay Validate Raw Material, Stability and Release Assay Evaluate Assay Performance Develop & Evaluate High Throughput Serology Assay Manufacturing Feasibility Studies Economic Feasibility Assessment Mfg.. Strategy Prepare Prelim. Eng.. & Basis of Design Inspection Preparation Prepare Lab Lots for POC Clinicals Prepare Pilot Lots for Dose Ranging Prepare Phase 3 Quality Lots Prepare Consistency Lots Prepare Launch Quantities of Vaccine Start Stage 0Stage 0 ReviewStage 1A ReviewStage 1B ReviewStage 2 ReviewStage 3 Review Dose Defined Regulatory Assessment Proof of Concept Bridging Studies Dose Ranging Studies Process Assessment- Clinicals Efficacy Studies Consistency Studies Scale Defined LPO Submit IND Prepare WMA Support WMA & Extend Expiry First Sale Release WMAApproval Product, Process & Formulation Finalized Kit Vendor Development of Serology Assays Capable of Evaluation Post-Vaccination and Post-Disease Seroconversion ID Parameter for POC Clinicals Stage 1 Anal.& Process Plan Spec.. Strategy & Rationale Validate In-Process and Product Characterization Assays Update Spec. Strategy & Rationale Transfer Assays Update Spec. Strategy & Rationale Update Spec. Strategy & Rationale Detail Design Order Equipment Build and Validate Facility Statement of Interest (SOI) RMC Approval Facilities Early Research Stability Experiments Preliminary Product & Packaging Definition Probe & Clinical Lot Stability Studies Pilot Lot Stability Studies Final Product & Primary Package Definition Post-Launch Product and Packaging Support Filling and Packaging Development Launch and Annual Stability Studies Full Scale Lot Stability Studies Market Container Stability Studies Final Secondary Package Definition

17. Physical Plant Design starts early, at risk Construction starts in early phase II Dig-to-validation from 3 to 5+ years Purpose-built to make one vaccine Back-up facilities are rare Accidents, or regulatory action can stop supply Raw material shortages, recalls Filling and packaging also fragile

18. Overall Timelines Inception to licensure can span a career Varicella, 29 years (1966-1995) MMRV, 23 years (1982-2005) Shingles, 21 years (1985-2006) RotaTeq®, 16 years (1990-2006 +) Gardasil®, 13 years (1993-2006) Persistence in a necessary element!