牛初乳免疫球蛋白M的分离纯化及多克隆抗体制备OA

This study aims to isolate and purify immunoglobulin M(IgM)from bovine colostrum and uses it as an antigen to immunize BALB/c mice for the preparation of mouse anti-bovine IgM polyclonal antibodies.After centrifugal degreasing and salting-out of bovine colostrum,IgM is isolated and purified by a combination of Sephacryl S-300 gel chromatography and DEAE-Sepharose 4FF ion-exchange chromatography.The obtained high-purity IgM is identified by SDS-PAGE.Subsequently,the purified IgM is used as an antigen to prepare mouse anti-bovine IgM polyclonal antibodies via immunization,and the titer and specificity of the antibodies are determined by ELISA,Western blot,and other methods.The results show that the molecular weights of the heavy and light chains of the isolated and purified IgM are 87.977±0.997 kDa and 25.952±0.160 kDa,respectively,with a purity of over 94%.The titer of the polyclonal antibody is 1∶512 000 as determined by ELISA,and a single specific band is observed in Western blot,indicating that the polyclonal antibody specifically reacts with bovine IgM and has good specificity.This study successfully isolates and purifies high-purity IgM from bovine colostrum and obtains mouse anti-bovine IgM polyclonal antibodies with high titer.