The Front-End of Vaccine Manufacturing: Getting Good Candidates from the Get-Go

1. The Front-End of Vaccine Manufacturing: Getting Good Candidates from the Get-Go William Warren, Donald Drake, Janice Moser, Haifeng Song, Eric Mishkin VaxDesign Corporation Orlando, FL 32826 www.vaxdesign.com Eric Eisenstadt, Hervé Tettelin, Scott Peterson The Institute For Genomic Research Rockville, MD 20850 www.tigr.org VaxDesign’s work was funded by DARPA/DSO in the Rapid Vaccine Assessment Program TIGR’s work was funded by NIH/NIAID & Novartis

2. High risk • Large investment • Ill-afford lost opportunity costs Manufacturing Facilities Begin During Clinical Trials http://www.vaccinealliance.org/site_repository/resources/21VacMarket.pdf

3. 5 – 15 years Conventional Vaccine Development DNA recombinant technology

4. Costs and Time Associated with Today’s Vaccine Product Lifetime Cycle • Challenges: • Can we obtain possible vaccine candidates faster? • Can we reduce the time to get vaccines to the marketplace? • Can we reduce the associated costs? • Can we make a more predictive and representative readout? • Can we have greater success in clinical trials?

5. Reverse Vaccinology: Applying Genomics, Immunology & Engineering To Rapidly Assess Vaccine Candidates High Throughput Gene Expression ~ 6 months - 1 year

6. Genomics, Tissue Engineering, & Automation Provide a New Approach • Genomics analysis of DNA sequence information identifies vaccine candidates that can be used alone or in combination • Tissue engineering provides direct access to predictive human immune response without using people • High throughput automation for repeatable, reproducible and rapid processes

7. The Systems Vaccinology Pipeline

8. Whole genome sequence In silico comparisons and antigen predictions Steps 1 & 2: Produce the genome sequence, read it, and predict the vaccine candidates (reverse vaccinology) TIGR: rapid sequencing technologies that have allowed us to clone thousands of open reading frames derived from the genomes of a variety of infectious agents, including influenza virus

9. Proof of Principle for Reverse Vaccinology via TIGR/Chiron Partnership Serogroup B Neisseria meningitidis - MenB No vaccine candidate in 40 years of classical vaccinology Genome sequence 7 novel candidates Antigenic, Accessible, Highly Conserved Specific and Bactericidal Tettelin et al. (2000) Science 287, 1809-1815 Pizza et al. (2000) Science 287, 1816-1820 Group B Streptococcus - GBS One genome sequenced - No candidate providing broad protection Tettelin et al. (2002) PNAS 99, 12391-12396 Analysis of 8 genomes Highly diverse species Cocktail of 4 candidates confer broad protection Tettelin et al. (2005) PNAS 102, 13950-13955 Maione et al. (2005) Science 309, 148-150

10. The Systems Vaccinology Pipeline

11. Step 3: Making the Vaccine Antigens viaHigh Throughput Expression • Directly from the pathogen genome via high-throughput technology that clones and translates the gene • Indirectly by synthesizing the gene de novo and then translate it

12. The Gateway Cloning Platform

13. Men B Vaccine: Genomic Approach Bottleneck and relevance? http://www.meningitis.org/uploads/C05_2_15_Rappuoli.pdf

14. The Systems Vaccinology Pipeline • Clinical trial in a test tube: high throughput in vitro assay system

15. Step 4: High-throughput testing of proteins as possible human vaccine candidates • ex vivo models of human immunity that are functionally equivalent to the human immune system • Meld immunology with engineering to find elegant, practical solutions to complex biological problems

16. Lymphoid Tissue Equivalent Module Vaccination Site Collagen Module Artificial Immune System Cell Interactions

17. How To Create a Functional Ex Vivo AIS Lymphoid Tissue Equivalent (LTE) Vaccination Site (VS) DC crossing endothelium Microbes and Infection (2003) 5: 205-212

18. Example: Representative Ex-Vivo Immunogenicity Testing Donor had a high anti-tetanus toxoid titer; yet, the industry standard PBMC assay failed to show protection

19. The artificial immune system construct supports the induction of naïve and recall human B cell responses

20. Predictive ex vivo Clinical Research For Influenza • Representative high-throughput ex vivo clinical research model that can assess initiation through neutralization immune responses of influenza/pandemic vaccine candidates • Rapid, predictive influenza/pandemic strain selection • Test immunity to circulating strains • Vaccine selection • Strains in which there are deficiencies or inappropriate responses HA-FITC Humoral VS (DCs) LTE (T/B) Neutralizing Ab Cellular

21. The Systems Vaccinology Pipeline Develop vaccines or fully human therapeutic mAbs

22. When Thinking of Vaccine Manufacturing …. • Companion diagnostics to better design clinical trials • e.g., Herceptin: only donors with Her2 receptors respond • HBV works on 80-90% of population • Couple in vitro culture techniques with rapid sequencing and expression technologies to create an automatable, high-throughput system for assessing clinical viral isolates to elicit specific immunity in the population at large

23. When Thinking of Vaccine Manufacturing …. • In-line immunogenicity with new manufacturing processes • e.g., Eprex® EPO induced immunity to EPO in some patients, which caused severe anemia • e.g., Biogenerics • e.g., New formulations • Generate wholly human mAbs • Use the AIS as an Ab biofactory

24. Need for New Predictive & Representative Vaccine Candidates Earlier in the Vaccine Development Pipeline • $51B spent for drug and vaccine discovery and development in 1995 • Increases by 7% each year • $1B in R&D cost for each new drug and vaccine approved, including failures • Predicted to reach $2B by 2010 • Manufacturing is an intimate part of these costs • Reverse Vaccinology may reduce costs to bring drugs to the market Reverse vaccinology http://www.bio-itworld.com/issues/2006/sept/2-billion-pill