Purification and characterization of MAP30 by immunoaffinity chromatography
Objective:To establish an immunoaffinity chromatography method for rapid purification of MAP30 polypeptide (a momordica toxin).Methods:New Zealand white rabbits (male) were immunized with the toxin MAP30 as an antigen to prepare antiserum. Polyclonal antibodies were purified by Mabselect affinity chromatography. Synthesized antibody-Sepharose 2B affinity chromatography medium using Sepharose 2B as a matrix coupled with polyclonal antibody was used to purify the momordica toxin MAP30.Results:The MAP30 toxin purified with the self-made immunoaffinity medium showed a single color band under reducing conditions on SDS-PAGE, and the corresponding relative molecular weights were all 30 kDa. The total sugar content measured by the sulfate-anthrone method was 1.54%. PC12 cells were used as the test cell type, and the maximum inhibition rate of MAP30 (anti-tumor activity) was 90%.Conclusion:The structure, properties, and activity characteristics of the purified target protein-MAP30 in this study are consistent with those reported in the literature, which suggests that the immunoaffinity chromatography method established in this study is feasible for the separation and purification of MAP30.
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