摘要
应用ＲＴＰＣＲ技术，从分泌抗人肝癌单克隆抗体（ｍＡｂ）的杂交瘤细胞ＨＡｂ１８中，扩增出抗体ＶＨ和ＶＬ基因，并测序。计算机分析表明，ＶＨ和ＶＬ均符合小鼠抗体可变区的特征，为功能性重排的抗体可变区基因。用（Ｇｌｙ４Ｓｅｒ）３连接肽基因，将ＶＨ和ＶＬ连接成ＳｃＦｖ基因，并进行序列测定，结果表明，ＶＨ，ＶＬ，ｌｉｎｋｅｒ拼接正确，基因全长为７２６ｂｐ。用噬菌粒表达载体ｐＣＡＮＴＡＢ５Ｅ在大肠杆菌中表达了可溶性的ＳｃＦｖ融合蛋白，经流式细胞仪分析表达产物可特异地与肝癌细胞结合。
The V H and V L genes were amplified from a hybridoma cell line HAb18 producing mouse mAb against human liver cancer by RT PCR. The genes were sequenced and compared with those published genes in EMBL Genbank. The results showed that the V H and V L genes were homologous with the published gene sequences of mouse antibody variable region. The V H and V L genes were connected through a flexible linker(Gly 4Ser) 3, and the V H linker V L(ScFv) fusion gene was cloned into a phagemid vector pCANTAB 5E and expressed in E.coli HB2151. Immunofluorescence assay showed that the expressed protein could bind to human liver cancer cell line SMMC 7721, but not to normal liver cell line QZG and gastric cancer cell line SCG 7901.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
1998年第2期102-104,109,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家863计划资助
