Study On The Purification And Characterization Of Ig-Binding Protein(IGBP) On Spleen Cell Membrane Of Common Carp(Cyprinus Carpio L.)

Serum IgM was purified from common carp(Cyprinus carpio L.) and then the IgM was used to make a affinity chromatography to purify the IgM binding protein(IGBP) on spleen cell membrane of common carp, and study the express of IGBP in spleen, intestine, skin and liver. Purified IgM was purified by polycarylamide gel electrophoresis(SDS-PAGE).The results indicated that the serum IgM can be separated into two subunits: the the heavy chain and the light chain,with the molecular masses were 82kDa and 29kDa, respectively.The spleen, intestine, skin and liver tissue were stained with different fluorescence marker. Firstly, the spleen, intestine, skin and liver tissue was labeled by FITC-IgM, and observed by confocal microscope, the results indicated that all above tissue were positive. Secondly, IGBP purified by affinity chromatography was used to prepare Guinea Pig antisera(Anti-IGBP) and then used to label the spleen, intestine, skin and liver tissue indirectly, the results showed that the IGBP was all expressed in spleen, intestine, skin and liver. The anti-IGBP was instead by normal Guinea Pig serum in the control group, and all the control group were negative. All the results indicated that there are molecular which can bind fish IgM.In this study, we find some evidences that there are IgM binding protein(IGBP) in the cells of spleen, intestine, skin and liver. It may involved in mucosal Ig transportation and other immune response in the adaptive immunity. So it would help to understand the mechanism of fish immunity.The purified IgM was used to make a chromatography for affinity purification of the IgM binding protein(IGBP) on spleen cell membrane. SDS-PAGE analysis showed that we have gotten three kinds of protein which could bind carp fish IgM.