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.
出处
《中华神经外科杂志》
CSCD
北大核心
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
关键词
脑肿瘤
胶质瘤
单克隆抗体
单链抗体
构建
Glioma Single chain antibody Gene recombination Prokaryotic expression
