1. November 14, 2007VRBPAC meeting1 of 24 Laboratory of Method Development Division of Viral Products Office of Vaccines Research and Review

2. November 14, 2007VRBPAC meeting2 of 24 LMD Mission Statement To conduct research leading to new concepts and methods for quality control and evaluation of existing and new viral vaccines and other biologicals

3. November 14, 2007VRBPAC meeting3 of 24 LMD Research Programs Chumakov: Evaluation of safety and potency of viral vaccines based on analysis of molecular consistency (Z01 BK 08013-03) Chizhikov: Microarray-Based Evaluation of Purity and Safety of Biological Products (Z01 BK 08014-03) Rubin: Developing tests to evaluate virus vaccine safety for the nervous system (Z01 BK 02023-03)

4. November 14, 2007VRBPAC meeting4 of 24 LMD Staff

5. November 14, 2007VRBPAC meeting5 of 24 Scope of Regulatory Responsibilities Poliovirus vaccine and combinations Mumps, Measles, Rubella, (Varicella) Lot release of IPV, MMR, Gardasil® QC of vaccines under licensure IND and BLA related to new viral vaccines

6. November 14, 2007VRBPAC meeting6 of 24 International Activities Advising WHO on vaccines issues International guidances / recommendations Validation of reference materials International Collaborative studies Biotechnology Engagement Program

7. November 14, 2007VRBPAC meeting7 of 24 Neuropathogenesis Team Tahir Malik, Ph.D., Staff Scientist Christian Sauder, PhD., Visiting Associate Cheryl Zhang, PhD, ORISE Fellow Malen Link, PhD, ORISE Fellow Laura Nerret, B.S., ORISE Fellow Steven Rubin, M.Sc. Acting Principal Investigator

8. November 14, 2007VRBPAC meeting8 of 24 Neuropathogenesis Team Overview Performs research on viral vaccines associated with adverse events in the central nervous system –Pre-clinical neurovirulence safety assays Mumps virus vaccines Influenza virus vaccines Vaccinia virus-based smallpox vaccines –Molecular basis of virus neurovirulence Mumps virus Influenza virus Serves as a unique neuropathogenesis resource for CBER Since the team represents one of only two active mumps virus research programs in the country, it serves as the agency’s expert on issues of mumps vaccine safety and efficacy

9. November 14, 2007VRBPAC meeting9 of 24 Recent Research Accomplishments Mumps Virus –Leading a WHO effort to perform a large scale formal validation effort to support use of a rat-based test, developed in our laboratory, for assessing the neurological safety of new candidate mumps virus vaccines (to replace the monkey neurovirulence safety test) –Established a primary neuronal cell culture system capable of differentiating neurotoxic from non-neurotoxic mumps virus strains –Developed infectious mumps virus cDNA clones for evaluating the role of specific virus genes/proteins in mumps virus neurotropism and neurotoxicity. –Established a standardized mumps virus neutralization assay for evaluation of mumps immunity with suggested surrogate endpoints for protection –Participating member of an interagency investigation of the 2006 mumps virus epidemic and provided key data demonstrating continued vaccine efficacy and data supporting an update to the ACIP recommendations for the control and elimination of mumps to include two doses of mumps-containing vaccine instead of one for documentation of immunity

10. November 14, 2007VRBPAC meeting10 of 24 Recent Research Accomplishments Influenza Virus –Developed a rat-based test for assessing the neurological safety of new candidate influenza virus vaccines –Established a plasmid DNA transfection system for the rescue of infectious influenza A virus to aid in the study of virus neurotropism and neurotoxicity Vaccinia Virus –Developed a rat-based test for assessing the neurological safety of new candidate vaccinia virus-based smallpox vaccines

11. November 14, 2007VRBPAC meeting11 of 24 Microarray team Dmitriy Volokhov, PhDPostdoctoral Fellow Hyesuk Kong, PhDPostdoctoral Fellow Vladimir Chizhikov, PhD Principal Investigator

12. November 14, 2007VRBPAC meeting12 of 24 MICROARRAY-BASED EVALUATION OF PURITY AND SAFETY OF BIOLOGICAL PRODUCTS Development and Improvement of Microarray Methods –Genotyping of a number of human and animal pathogens Product Safety and Purity –Detection of extraneous agents (mycoplasma)

13. November 14, 2007VRBPAC meeting13 of 24 Development and Improvement of Microarray Methods Advanced microarray substrates and protocols Not-fluorescent detection (nano-gold particles) Lab-on-Chip (collaboration with CDRH) Drug-resistance of M. tuberculosis (in collaboration with JHU) Genotyping of measles isolates (in collaboration with JHU) Genotyping of VZV isolates (in collaboration with CDC) P and G genotyping of rotaviruses (in collaboration with NIAID) Detection of bioterrorism pathogens in blood (in collaboration with OBRR) Detection and analysis of food-borne pathogens (in collaboration with CDRH, CFSAN) Technical approachesBiological models

14. November 14, 2007VRBPAC meeting14 of 24 Main Accomplishments (2003-2007): Establishment of FDA collection of mycoplasmas (>200 species) Genetic analysis of the 16S-23S Intergenic spaces, potential marker for identification of mycoplasma species Development and evaluation of a PCR/microarray assay based on nano-gold particle technology for detection identification of Mollicutes species. Development of a cell culture-based method for biological enrichment of mycoplasma contaminants that allows for significant increase of the sensitivity of mycoplasma detection and three-fold reduction of the mycoplasma testing time. Development and evaluation of novel microarrays for detection and analysis of important human pathogens and their virulence factors (genotyping of Clostridium perfringens toxins, screening new mutations in the structural region West Nile virus genome, identification of G and P genotypes of rotaviruses, analysis of multiple staphylococcal enterotoxin genes, and detection and analysis of bioterrorism pathogens in blood et al.)

15. November 14, 2007VRBPAC meeting15 of 24 Productivity (2003-2007): 2 patents (together with ATCC and OBRR/FDA) 24 publications in peer-reviewed scientific journals 2 manuscripts in preparation 4 invited talks including 1st International Mycosafe Symposium, April 2007 (Austria) Several poster presentations on different international, national and FDA scientific meetings

16. November 14, 2007VRBPAC meeting16 of 24 Molecular consistency of viral vaccines Eugenia Dragunsky, MD, PhDStaff Scientist Gennady Rezapkin, PhDStaff Scientist Majid Laassri, PhDVisiting Associate Joan Enterline, BABiologist Monica Parker, BSBiologist Alexander Neverov, PhDPostdoctoral Fellow Alexander Ivanov, MD, PhD, DSciPostdoctoral Fellow (resigned 11-07) Elena Cherkasova, PhDPostdoctoral Fellow (resigned 05-07) Konstantin Chumakov, PhD Principal investigator

17. November 14, 2007VRBPAC meeting17 of 24 Principal directions Oral polio vaccine Inactivated polio vaccine Flavivirus recombinant vaccines Influenza vaccine Microarray methods for genotyping of viruses (Orthopox, Herpes viruses) Molecular consistency of cell substrates (Vero cells)

18. November 14, 2007VRBPAC meeting18 of 24 Oral poliovirus vaccine Tg mouse test for neurovirulence of OPV Molecular basis of neurovirulence and interactions with host immune system Genetic stability of OPV –In vitro studies –Clinical studies Evolution of vaccine-derived polioviruses –Genomic evolution –Antigenic drift

19. November 14, 2007VRBPAC meeting19 of 24 Advanced Immunochemical Methods Block-ELISA procedure Epitope profiling Paratope profiling

20. November 14, 2007VRBPAC meeting20 of 24 Inactivated Polio Vaccine Evaluation of Sabin IPV Combination vaccines Transgenic mouse immunization / challenge protection test Effect of novel adjuvants Alternative routes of administration

21. November 14, 2007VRBPAC meeting21 of 24 Flavivirus vaccines West Nile / Dengue 4 chimeras TBE (Langat) / Dengue 4 chimeras Microarray analysis of genetic stability

22. November 14, 2007VRBPAC meeting22 of 24 Seasonal Influenza Vaccines Microarray method for rapid genotyping of genomic segments of reassortant vaccine strains –Influenza B –Influenza A

23. November 14, 2007VRBPAC meeting23 of 24 Genotyping of Orthopox and Herpes viruses Microarray - based method DARPA grant BTEP-sponsored collaboration

24. November 14, 2007VRBPAC meeting24 of 24 Molecular Consistency of Cell Substrates Tumorigenicity of Vero cells Surrogate markers of tumorigenicity –mitochondrial DNA analysis –microarray gene expression data –proteomics Markers of cell culture confluence –microarray analysis