Application of deglycosylation to SDS PAGE analysis improves calibration of influenza antigen standards.

Abstract

Each year the production of seasonal influenza vaccines requires antigen standards to be available for the potency assessment of vaccine batches. These are calibrated and assigned a value for haemagglutinin (HA) content. The calibration of an antigen standard is carried out in a collaborative study amongst a small number of national regulatory laboratories which are designated by WHO as Essential Regulatory Laboratories (ERLs) for the purposes of influenza vaccine standardisation. The calibration involves two steps; first the determination of HA protein in a primary liquid standard by measurement of total protein in a purified influenza virus preparation followed by determination of the proportion of HA as determined by PAGE analysis of the sample; and second, the calibration of the freeze-dried reference antigen against the primary standard by single radial immunodiffusion (SRD) assay. Here we describe a collaborative study to assess the effect of adding a deglycosylation step prior to the SDS-PAGE analysis for the assessment of relative HA content. We found that while the final agreed HA value of the samples tested was not significantly different with or without deglycosylation, the deglycosylation step greatly improved between-laboratory agreement.

• Authors: Ruth Harvey, Michelle Hamill, James S Robertson, Philip D Minor, Galina M Vodeiko, Jerry P Weir, Hitoshi Takahashi, Yuichi Harada, Shigeyuki Itamura, Pearl Bamford, Tania Dalla Pozza, Othmar G Engelhardt
• Journal: Biologicals : journal of the International Association of Biological Standardization (Biologicals) Vol. 40 Issue 1 Pg. 96-9 (Jan 2012) ISSN: 1095-8320 [Electronic] England
• PMID: 22244521 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 The International Alliance for Biological Standardization. All rights reserved.

Chemical References

• Antigens, Viral
• H1N1 virus hemagglutinin
• Hemagglutinin Glycoproteins, Influenza Virus
• Influenza Vaccines

Topics

• Antigens, Viral (chemistry)
• Calibration
• Electrophoresis, Polyacrylamide Gel (methods, standards)
• Glycosylation
• Hemagglutinin Glycoproteins, Influenza Virus (chemistry)
• Influenza A Virus, H1N1 Subtype (chemistry)
• Influenza A Virus, H3N2 Subtype (chemistry)
• Influenza Vaccines (chemistry)
• World Health Organization