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
Eric Eisenstadt, Hervé Tettelin, Scott Peterson
The Institute For Genomic Research
Rockville, MD 20850
VaxDesign’s work was funded by DARPA/DSO in the
Rapid Vaccine Assessment Program
TIGR’s work was funded by NIH/NIAID & Novartis

2. Manufacturing Facilities Begin During Clinical Trials
High risk
Large investment
Ill-afford lost opportunity costs

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

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. and antigen predictions
Steps 1 & 2: Produce the genome sequence, read it, and predict the vaccine candidates (reverse vaccinology)
In silico comparisons
and antigen predictions
Whole genome sequence
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,
Pizza et al. (2000) Science 287,
Group B Streptococcus - GBS
One genome sequenced - No candidate providing broad protection
Tettelin et al. (2002) PNAS 99,
Analysis of 8 genomes
Highly diverse species
Cocktail of 4 candidates confer broad protection
Tettelin et al. (2005) PNAS 102,
Maione et al. (2005) Science 309,

10. The Systems Vaccinology Pipeline

11. Step 3: Making the Vaccine Antigens via High 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?

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

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

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 VS
(DCs)
LTE
(T/B)
Humoral
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