title: Development and applications of universal H7 subtype-specific antibodies for the analysis of influenza H7N9 vaccines.
abstract: H7N9 is a newly emerged avian influenza virus with a relatively high mortality rate in humans. At this time, there is no licensed vaccine for human protection. Development of analytical tools for H7N9 vaccine could facilitate vaccine development. Here, a universally conserved epitope in all H7 hemagglutinin (HA) sequences was identified through comprehensive bioinformatics analyses. The peptide epitope, RSGSSFYAEMK, (aa positions 149 to 159), is located on the head of the HA molecule. Antibodies generated against this universal H7 epitope were remarkably specific against H7 viral sequence with no detectable cross-reactivity to other HA subtypes. A new immunoblotting assay based on the universal H7 antibody was developed and compared with the traditional single radial immunodiffusion assay (SRID) for potency analyses of candidate H7N9 vaccines. This new assay was more sensitive and rapid compared to SRID. In addition to statistically acceptable precision and reproducibility, the new assay differs from many other alternative potency assays for influenza vaccine in that it is potentially stability-indicating, which is an important requirement for industry vaccine stability studies analyses. Furthermore, the robustness of this new assay was demonstrated by the quantitative determination of HA content in four H7N9 vaccines (split or inactivated) from different manufacturers.
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Ottawa, ON, Canada.
authors.forename: Caroline
authors.initials: C
authors.lastname: Gravel
authors.affiliation: Canadian Food Inspection Agency, Ottawa, ON, Canada.
authors.forename: Cathie
authors.initials: C
authors.lastname: Elmgren
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Ottawa, ON, Canada; Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON, Canada.
authors.forename: Abenaya
authors.initials: A
authors.lastname: Muralidharan
authors.affiliation: Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia; Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Anwar M
authors.initials: AM
authors.lastname: Hashem
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Ottawa, ON, Canada.
authors.forename: Bozena
authors.initials: B
authors.lastname: Jaentschke
authors.affiliation: National Institutes for the Control of Food and Drug and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Beijing, PR China.
authors.forename: Kangwei
authors.initials: K
authors.lastname: Xu
authors.affiliation: Canadian Food Inspection Agency, Ottawa, ON, Canada.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Widdison
authors.affiliation: Canadian Food Inspection Agency, Ottawa, ON, Canada.
authors.forename: Kristin
authors.initials: K
authors.lastname: Arnold
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Ottawa, ON, Canada.
authors.forename: Aaron
authors.initials: A
authors.lastname: Farnsworth
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Ottawa, ON, Canada.
authors.forename: Aline
authors.initials: A
authors.lastname: Rinfret
authors.affiliation: National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada.
authors.forename: Gary
authors.initials: G
authors.lastname: Van Domselaar
authors.affiliation: National Institutes for the Control of Food and Drug and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Beijing, PR China.
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
authors.affiliation: National Institutes for the Control of Food and Drug and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Beijing, PR China.
authors.forename: Changgui
authors.initials: C
authors.lastname: Li
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Ottawa, ON, Canada; Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON, Canada. Electronic address: sean.li@hc-sc.gc.ca.
authors.forename: Xuguang
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D015151:Immunoblotting / Q000379:methods*; D005779:Immunodiffusion; D064766:Influenza A Virus, H7N9 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000276:immunology*; D051379:Mice; D011817:Rabbits; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D018985:Epitopes, B-Lymphocyte; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; C109963:hemagglutinin, avian influenza A virus
keywords: H7N9 influenza vaccine; Universal H7 epitope; Vaccine quantitation