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抗人卵巢癌×抗人CD3双特异性单链抗体的构建、表达及复性研究

Construction and Expression of an Anti-human Ovarian Carcinoma×Anti- human CD3 Bispecific Single-chain Antibody and its Refolding Studies

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【作者】 方敏赵瑞李骅蒋欣尹长城林晴黄华

【Author】 Fang Min, Zhao Rui, Li Hua, Jiang Xin, Yin Changcheng, Lin Qing, Huang Hualiang (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101)

【机构】 中国科学院遗传与发育生物学研究所西北农林科技大学生命科学院中国科学院遗传与发育生物学研究所 北京100101杨凌712100北京100101北京100101

【摘要】 构建了抗人卵巢癌×抗人CD3双特异性单链抗体 (scBsAb) ,在大肠杆菌中得到表达。用稀释复性 ,缓慢透析复性和凝胶过滤层析复性三种方法对scBsAb表达形成的包涵体进行复性研究表明 ,凝胶过滤层析复性能有效抑制蛋白的聚集。ELISA检测显示 ,复性后的scBsAb具有较高活性 ,能与相应抗原特异结合。

【Abstract】 An anti human ovarian carcinoma×anti human CD3 bispecific single chain antibody(scBsAb) is constructed and expressed in the E. coli strain GI724 as inclusion body. Three different refolding procedures had been sused to perform renaturation of the scBsAb. Direct dilution and gradual dialysis both leaded to aggregation of the refolded proteins. However, size exclusion chromatography performing reneturaton could supprss protein aggregation. ELISA tests demonstrate that the refolded scBsAb has good activity, can bind to its target antigens with specificity.

【基金】 863计划 (863 10 2 0 9 0 4 0 1)资助项目
  • 【文献出处】 高技术通讯 ,High Technology Letters , 编辑部邮箱 ,2002年11期
  • 【分类号】R737.31
  • 【被引频次】6
  • 【下载频次】109
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