抗人脑胶质瘤单克隆抗体SZ39单链抗体的构建及初步表达  

The construction and expression of single chain antibody SZ39 ScFv for human brain glioma

作　　者：沈树泉[1,2] 周筱梅[1,2] 黄强[1,2] 许志元[1,2] 王爱东[1,2] 杜子威[1,2] 

机构地区：[1]苏州医学院附属二院神经外科脑肿瘤研究室 [2]上海市肿瘤研究所

出　　处：《中华神经外科杂志》1997年第5期260-262,共3页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金;工业基金

摘　　要：目的：研究人脑胶质瘤基因工程单抗。方法：采用基因重组技术分别将抗人脑胶质瘤单抗ＳＺ３９重链和轻链可变区基因片段连接，构建表达载体，并在大肠杆菌中表达。结果：表达的ＳｃＦｖ抗体为可溶性，ＥＬＩＳＡ和ＷｅｓｔｅｒｎＢｌｏｔ检测证实能特异性地与人脑胶质瘤细胞系ＳＨＧ－４４表面抗原结合，其表达量为２２０μｇ／Ｌ。结论：成功地构建了抗人脑胶质瘤单抗ＳＺ３９ＳｃＦｖ表达质粒，并得到了与胶质瘤细胞表面抗原相结合的单链抗体。Objective: To study the genetically engineered single chain antibody of human brain glioma. Methods: The single chain antibody SZ39 ScFv specific for human brain glioma was produced by recombinant DNA techniques. The variable region gene fragments of SZ39 heavy and light chain which were cloned from SZ 39 hybridoma cells were inserted into pHEN1 expression vectors containing universal linker gene, respectively. The construct was transformed into E. Coli HB2151 and ScFv was expressed and secreted into supernatant of culture medium. Results: The expressed ScFv antibody was soluble, and its specific bining capacity to the surface associated antigen of human glioma cell line SHG 44 was further comfirmed with ELISA and Western Blot assay. The yield of ScFv secreted into the culture medium was 220μg/L. Conclusion: We have successfully constructed the expression vector of ScFv of the anti human glioma monoclonal antibody SZ39, which can be expressed in E. Coli and showed high, specific affinity for the glioma cell surface associated antigen.

关 键 词：脑肿瘤 胶质瘤 单克隆抗体 单链抗体 构建 

分 类 号：R392－33[医药卫生—免疫学] R739.410.5[医药卫生—基础医学]