A hemagglutinin quantification method for development of an influenza pandemic vaccine using size exclusion high performance liquid chromatography.

Abstract

Single radial immunodiffusion (SRID) assay requires a reference antigen and an antibody to the hemagglutinin (HA) of an influenza vaccine. As it takes 2‑3 months to develop the reference antigen, vaccine development is delayed in cases of an influenza pandemic. In the present study, the measurement of the HA content of influenza vaccines was assessed using size exclusion high performance liquid chromatography (SE‑HPLC) for the rapid development of a pandemic vaccine. When the 2009 H1N1 reference antigen, pandemic 2009 H1N1 vaccine and 2010 seasonal influenza vaccines were analyzed by SE‑HPLC, the HA of the reference antigen and vaccines was specifically separated. The presence and specificity of HA were evidenced with immunoprecipitation and ELISA assays. For the influenza vaccines, the chromatogram pattern and retention time of HA were similar among the antigen types (2009 H1N1, 2010 H3N2 and 2010 B). In addition, when SE‑HPLC was applied, the ratio of HA chromatogram to peak area revealed a significant correlation with HA concentration for the reference antigen and vaccine. The result of the HA content calculation based on SE‑HPLC exhibited 99.91‑100% similarity, compared with that of SRID. These findings suggest that the measurement of peak area ratio/HA content using SE‑HPLC may be a substitute for SRID and rapidly measure HA content to enable faster development of a vaccine during an influenza pandemic.

• Authors: Hang Sik Roh, Hye Min Song, Bo Reum Yun, Hyun Kyung Kang, Keum Suk Choi, Yun Ju Park, Dong Sub Kim, Seung Hee Kim, In Pil Mo, Beum-Soo An, Chi Young Ahn
• Journal: Molecular medicine reports (Mol Med Rep) Vol. 11 Issue 4 Pg. 2819-24 (Apr 2015) ISSN: 1791-3004 [Electronic] Greece
• PMID: 25482872 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Hemagglutinins, Viral
• Influenza Vaccines

Topics

• Chromatography, High Pressure Liquid
• Hemagglutinins, Viral (chemistry, immunology)
• Humans
• Influenza A Virus, H1N1 Subtype (immunology)
• Influenza A virus (immunology)
• Influenza Vaccines (chemistry, immunology)
• Influenza, Human (prevention & control)