title: Application of deglycosylation and electrophoresis to the quantification of influenza viral hemagglutinins facilitating the production of 2009 pandemic influenza (H1N1) vaccines at multiple manufacturing sites in China.
abstract: The single radial immunodiffusion (SRID) method currently used to determine the hemagglutinin (HA) content of the inactivated influenza vaccines depends on the availability of reference HA antigen and corresponding anti-serum, updated and provided annually by World Health Organization (WHO) collaborative centers. Particularly early in a pandemic outbreak, reference reagents could be the bottleneck in vaccine development and release. Therefore, other reliable tests capable of quantifying HA content could substantially shorten the time needed for vaccine formulation. Here electrophoretic separation of deglycosylated samples in conjunction with densitometry was used to quantify HA contents of H1N1 vaccine at multiple manufacturing sites. We found the overall consistency between the alternative method and traditional SRID was 88-122% in seven lots of vaccine bulks from four subtypes (types) of influenza vaccine, confirming its suitability to quantify HA content. Moreover, we used the alternative method to prepare a national HA antigen reference in China for quality control of 2009 pandemic influenza A (H1N1) vaccines prior to the arrival of the WHO SRID reference standards, subsequently confirming good agreement between both methods. The alternative method for vaccine quantification enabled the Chinese health authority to approve H1N1 vaccine 1 month earlier than otherwise possible.
authors.affiliation: National Institute for the Control of Pharmaceutical and Biological Products, No. 2, Tiantan Xili, Beijing, 100050, PR China.
authors.forename: Changgui
authors.initials: C
authors.lastname: Li
authors.forename: Ming
authors.initials: M
authors.lastname: Shao
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Cui
authors.forename: Yingli
authors.initials: Y
authors.lastname: Song
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.forename: Liyong
authors.initials: L
authors.lastname: Yuan
authors.forename: Hanhua
authors.initials: H
authors.lastname: Fang
authors.forename: Zhenglun
authors.initials: Z
authors.lastname: Liang
authors.forename: Terry D
authors.initials: TD
authors.lastname: Cyr
authors.forename: Fengxiang
authors.initials: F
authors.lastname: Li
authors.forename: Xuguang
authors.initials: X
authors.lastname: Li
authors.forename: Junzhi
authors.initials: J
authors.lastname: Wang
mesh_terms: D002681:China; D004196:Disease Outbreaks / Q000266:history; D004591:Electrophoresis, Polyacrylamide Gel; D005843:Geography; D006031:Glycosylation; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis* / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D049674:History, 21st Century; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry / Q000592:standards; D007251:Influenza, Human / Q000453:epidemiology / Q000276:immunology / Q000517:prevention & control; D008420:Manufactured Materials / Q000032:analysis; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism* / Q000494:pharmacology; D011786:Quality Control; D012015:Reference Standards; D014944:World Health Organization
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase