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A new method for the preparation of anti-serum against influenza virus hemagglutinin for single radial immunodiffusion assay

GU Qin1,LI Juan2,XU Kang-wei2,et al.(1.Institute of Medical Biology,Chinese Academy of Medical Science,Kunming 650118,China;2.National Institutes for Food and Drug Control,Beijing 100050,China)  
Single Radial Immunodiffusion(SRID) is used for the determination of Hemagglutinin(HA) content in influenza vaccines.The standard antisera for SRID test are routinely provided by WHO collaborative laboratories.The traditional preparation method involves purification and cleavage of virus,further separating of HA fragments for immunization of animals.This whole process could take more than 3 months for completion,which delay the vaccine development in due course.In this study,the HA gene fragment was obtained by RT-PCR and cloned into bacterial expression vector.The recombinant proteins were purified and identified by SDS-PAGE and Western blot.After immunization of rabbits with re-naturation proteins,the antiserum was proved to be suitable for SRID test.The new method is helpful to a fast preparation of antiserum to be used for influenza vaccine SRID test.
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