Purification，Tag and Method for Detection of Grass Carp Serum IgM

Abstract

Abstract: Protein A affinity chromatography was used to purify immunoglobulin IgM from grass carp serum, and the purity and molecular weight of the product was analyzed by SDS-PAGE. Rabbit polyclonal antibodies of the purified grass carp IgM were prepared and detected by ELISA. The results showed that molecular weights of the light chain and heavy chain of the purified grass carp IgM were about 25 kD and 75 kD, respectively, and high titers of rabbit anti-grass carp IgM antiserum was obtained. Horseradish peroxidase was used to label the rabbit anti-grass carp IgM antibody. The labeled antibody was used to detect the antibody levels in serum of grass carp after immunizing by capsid protein VP4 and VP5 of grass carp hemorrhage virus isolated from Guangdong province（GCRV-GD108）. The results revealed that the labeled anti-grass carp IgM antibody could be used for the rapid detection of the IgM levels in grass carp serum. In this study, method to detect IgM levels in grass carp serum was established, which would provide a convenient method of serological testing for further study of pathogen invasion mechanism, grass carp immune defense mechanism and evaluation of grass carp vaccine effectiveness.

Key words: Immunoglobulin, Polyclonal antibodies ELISA, Horseradish peroxidase, GCRV-GD108 strain, Evaluation of vaccine effectiveness

Tian Yuanyuan, Ye Xing, Zhang Lili . Purification，Tag and Method for Detection of Grass Carp Serum IgM[J]. Biotechnology Bulletin, 2013, 0(4): 194-200.

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