重组人骨形态发生蛋白-2抗体的制备及纯化  

Preparation and Purification of Antibody for rhBMP-2

作　　者：赵丹慧[1] 张艳[1] 袁越[1] 李成文[1] 

机构地区：[1]北京积水潭医院北京市创伤骨科研究所,北京100035

出　　处：《中国生物工程杂志》2005年第7期45-49,共5页China Biotechnology

基　　金：北京市科委骨与关节疾病研究中心资助项目

摘　　要：骨形态发生蛋白-2是骨科研究领域的重要生物蛋白。用重组人骨形态发生蛋白-2(rhBMP2)与BSA连接成复合物作为免疫原,免疫家兔制备出多克隆抗血清。分别用免疫沉淀法和Sepharose4B-BSA反向免疫亲和吸附法去除抗血清中载体BSA抗体。用盐析法和蛋白层析法纯化出抗rhBMP2抗体。研究发现,家兔产生高效价抗体的最佳时间为14-16周,从rhBMP2抗体的特异性、相对纯度及回收率综合分析,免疫沉淀法去除BSA抗体优于反相亲和吸附法。Human bone morphogenetic proteins-2 (hBMP2 ) is an important protein in bone research area. rhBMP2 polyclone antiserum were produced by using rhBMP2-BSA as antigen. Two methods for removing carrier BSA antibody were compared between immunoprecipitation method and Sepharose 4B-BSA reversed-phase affinity immunosorbent method. The rhBMP2 antibody were purificated with saturated ammonium sulfate precipitation and protein A chromatography. The results show that the best time for producing high liter antibody of rhBMP2 in rabbits is about 14 - 16 weeks. Analyses reveal that the method of immunoprecipitation has more advantage than that of reversed-phase affinity immunosorbent for removing BSA antibody.

关 键 词：骨形态发生蛋白-2 重组人 抗体 纯化 制备 SEPHAROSE 免疫沉淀法 BSA 生物蛋白 研究领域 最佳时间 综合分析 抗血清 吸附法 免疫原 复合物 层析法 盐析法 特异性 回收率 家兔 去除 亲和 克隆 载体 

分 类 号：Q51[生物学—生物化学] S858.315.3[农业科学—临床兽医学]