title: An alternative method for preparation of pandemic influenza strain-specific antibody for vaccine potency determination.
abstract: The traditional assay used to measure potency of inactivated influenza vaccines is a single-radial immunodiffusion (SRID) assay that utilizes an influenza strain-specific antibody to measure the content of virus hemagglutinin (HA) in the vaccine in comparison to a homologous HA reference antigen. Since timely preparation of potency reagents by regulatory authorities is challenging and always a potential bottleneck in influenza vaccine production, it is extremely important that additional approaches for reagent development be available, particularly in the event of an emerging pandemic influenza virus. An alternative method for preparation of strain-specific antibody that can be used for SRID potency assay is described. The approach does not require the presence or purification of influenza virus, and furthermore, is not limited by the success of the traditional technique of bromelain digestion and purification of virus HA. Multiple mammalian expression vectors, including plasmid and modified vaccinia virus Ankara (MVA) vectors expressing the HAs of two H5N1 influenza viruses and the HA of the recently emerging pandemic H1N1 (2009) virus, were developed. An immunization scheme was designed for the sequential immunization of animals by direct vector injection followed by protein booster immunization using influenza HA produced in vitro from MVA vector infection of cells in culture. Each HA antibody was highly specific as shown by hemagglutination inhibition assay and the ability to serve as a capture antibody in ELISA. Importantly, each H5N1 antibody and the pandemic H1N1 (2009) antibody preparation were suitable for use in SRID assays for determining the potency of pandemic influenza virus vaccines. The results demonstrate a feasible approach for addressing one of the potential bottlenecks in inactivated pandemic influenza vaccine production and are particularly important in light of the difficulties in preparation of potency reagent antibody for pandemic H1N1 (2009) virus vaccines.
authors.affiliation: Division of Viral Products, Center for Biologics Evaluations and Research, Food and Drug Administration, 8800 Rockville Pike, Bethesda, MD 20892, United States.
authors.forename: Falko
authors.initials: F
authors.lastname: Schmeisser
authors.forename: Galina M
authors.initials: GM
authors.lastname: Vodeiko
authors.forename: Vladimir Y
authors.initials: VY
authors.lastname: Lugovtsev
authors.forename: Richard R
authors.initials: RR
authors.lastname: Stout
authors.forename: Jerry P
authors.initials: JP
authors.lastname: Weir
mesh_terms: D000818:Animals; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D006389:Hemagglutinins, Viral / Q000276:immunology; D006801:Humans; D005779:Immunodiffusion / Q000379:methods; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology; D007252:Influenza Vaccines / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D013678:Technology, Pharmaceutical / Q000379:methods*; D015164:Vaccines, Inactivated / Q000032:analysis / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D006389:Hemagglutinins, Viral; D007252:Influenza Vaccines; D015164:Vaccines, Inactivated; C458881:hemagglutinin fusogenic peptide, influenza virus