| 重组人骨形态发生蛋白-2抗体的制备及纯化 |
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| 引用本文： | 赵丹慧,张艳,袁越,李成文.重组人骨形态发生蛋白-2抗体的制备及纯化[J].中国生物工程杂志,2005,25(7):45-49. |
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| 作者姓名： | 赵丹慧 张艳 袁越 李成文 |
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| 作者单位： | 北京积水潭医院北京市创伤骨科研究所, 北京 100035 |
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| 基金项目： | 北京市科委骨与关节疾病研究中心资助项目 |
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| 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。
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| 关 键 词： | rhBMP2 多克隆抗体 纯化方法 |
| 收稿时间： | 2005-01-18 |
| 修稿时间： | 2005年1月18日 |
Preparation and Purification of Antibody for rhBMP-2 |
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| ZHAO Dan-hui,ZHANG Yan,YUAN Yue,LI Cheng-wen.Preparation and Purification of Antibody for rhBMP-2[J].China Biotechnology,2005,25(7):45-49. |
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| Authors: | ZHAO Dan-hui ZHANG Yan YUAN Yue LI Cheng-wen |
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| Abstract: | 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. |
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| Keywords: | rhBMP2 Polyclone antibodies Purification method |
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