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KMID : 0379520160320040269
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한국독성학회지
2016 Volume.32 No. 4 p.269 ~ p.274
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Evaluation of Haemagglutinin Content by RP-HPLC to Generate Pandemic Influenza Vaccine
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Kang Hyun-Kyung
Roh Hang-Sik
Song Hye-Min
Lee Kwang-Moon
Chung Seung-Tae
Ban Sang-Ja
Mo In-Pil
An Beum-Soo
Ahn Chi-Young
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Abstract
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The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.
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KEYWORD
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Influenza, Hemagglutinin, RP- HPLC, Vaccine
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