November 14, 2007VRBPAC meeting1 of 24 Laboratory of Method Development Division of Viral Products...

November 14, 2007 VRBPAC meeting 1 of 24

Laboratory of Method Development

Division of Viral Products

Office of Vaccines Research and Review


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


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)


Name Position NoteKonstantin Chumakov, PhD, DSci Laboratory Chief Principal Investigator

Eugenia Dragunsky, MD, PhD Staff ScientistGennady Rezapkin, PhD Staff ScientistMajid Laassri, PhD Visiting AssociateJoan Enterline, BA BiologistMonica Parker, BS BiologistAlexander Ivanov, MD, PhD, DSci Postdoctoral FellowElena Cherkasova, PhD Postdoctoral Fellow Resigned May 2007Alexander Neverov, PhD Postdoctoral Fellow Resigned November 2007Ronald Lundquist, PhD Microbiologist Regulatory work only

Vladimir Chizhikov, PhD Visiting Scientist Principal InvestigatorDmitriy Volokhov, PhD Postdoctoral FellowHyesuk Kong, PhD Postdoctoral Fellow

Kathryn M. Carbone, MD Team Leader Resigned November 2007Steven Rubin, MS Staff Scinetist Acting Principal Investigator

Tahir Malik, PhD Staff ScientistChristian Sauder, PhD Postdoctoral FellowQi Li, Ph.D Postdoctoral FellowCheryl Zhang, MD Postdoctoral FellowCandie Wolbert, MS Predoctoral FellowKari Vandenburgh, BS Predoctoral FellowMikhail Pletnikov, MD, PhD Guest Worker

LMD Staff


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


International Activities

• Advising WHO on vaccines issues

• International guidances / recommendations

• Validation of reference materials

• International Collaborative studies

• Biotechnology Engagement Program


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


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


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


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


Microarray team

• Dmitriy Volokhov, PhD Postdoctoral Fellow• Hyesuk Kong, PhD Postdoctoral Fellow

Vladimir Chizhikov, PhD

Principal Investigator


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)


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 approaches Biological models


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.)


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


Molecular consistency of viral vaccines

• Eugenia Dragunsky, MD, PhD Staff Scientist• Gennady Rezapkin, PhD Staff Scientist• Majid Laassri, PhD Visiting Associate• Joan Enterline, BA Biologist• Monica Parker, BS Biologist• Alexander Neverov, PhD Postdoctoral Fellow • Alexander Ivanov, MD, PhD, DSci Postdoctoral Fellow (resigned 11-07)• Elena Cherkasova, PhD Postdoctoral Fellow (resigned 05-07)

Konstantin Chumakov, PhD

Principal investigator


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)


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


Advanced Immunochemical Methods

• Block-ELISA procedure

• Epitope profiling

• Paratope profiling


Inactivated Polio Vaccine

• Evaluation of Sabin IPV

• Combination vaccines

• Transgenic mouse immunization / challenge protection test

• Effect of novel adjuvants

• Alternative routes of administration


Flavivirus vaccines

• West Nile / Dengue 4 chimeras

• TBE (Langat) / Dengue 4 chimeras

• Microarray analysis of genetic stability


Seasonal Influenza Vaccines

• Microarray method for rapid genotyping of genomic segments of reassortant vaccine strains– Influenza B– Influenza A


Genotyping of Orthopox and Herpes viruses

• Microarray - based method

• DARPA grant

• BTEP-sponsored collaboration


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

November 14, 2007VRBPAC meeting1 of 24 Laboratory of Method Development Division of Viral Products...

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