冉宇靓 杨治华 王贵齐 刘军 孙立新 董志伟 100021 北京，中国医学科学院中国协和医科大学肿瘤研究所肿瘤医院 中华肿瘤杂志 1999 0 21 6


    关键词：血管内皮细胞生长因子；嵌合抗体；真核细胞表达 期刊 zhzlzz 0 基础研究 fur -->


    

【摘要】 目的 在真核细胞中表达抗人血管内皮生长因子165(vascularendothelial growth factor 165, VEGF165)人/鼠嵌合抗体。方法 将抗VEGF165鼠单抗VmD11的轻、重链可变区基因克隆入基因工程抗体真核表达载体中，转染二氢叶酸还原酶缺陷型中国仓鼠卵巢(dihydrofolate reductase-deficient Chinese hamster ovary, CHO-dhfr^- )细胞进行表达，采用RT-PCR、ELISA、Westernblot等技术检测表达产物的人源性与抗原结合特异性。结果 从转染及筛选后的CHO细胞培养上清中，检测到含有人抗体恒定区Cγ1和Cκ的抗VEGF165人/鼠嵌合抗体，RT-PCR也证实了该嵌合抗体在mRNA水平上的表达。结论 在真核细胞中表达了具有VEGF165抗原特异性的人/鼠嵌合抗体。

Construction ofanti-human VEGF165 chimeric antibodies and expression in eukaryotic cells

RAN Yuliang, YANG Zhihua, WANG Guiqi, et al. Cancer Institute(Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing100021

【Abstract】 Objective To express chimericanti-VEGF165 antilody genes in eukaryotic cells.MethodsThe variable region genes of lightand heavy chains from mouse anti-human VEGF165 monoclonal antibody VmD11 were cloned intoeukaryotic expression vectors and transfected into dihydrofolate reductase-deficientChinese hamster ovary(CHO-dhfr^- ) cells to express chimeric antibody. Theantibody expressed was examined for the presence of human constant regions and specificityagainst human VEGF165.Results The chimeric antibody with human constant regions andspecificity against human VEGF was detected in the culture supernatant of transfected CHOcells by ELISA and Western blot. The chimeric antibody gene was also detected at mRNAlevel by RT-PCR.Conclusion The mouse anti-human VEGF165 monoclonal antibody issuccessfully humanized and expressed in eukaryotic cells.

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