hTNF-α单克隆抗体轻、重链可变区基因的克隆和序列及分子结构分析  

Cloning, Sequence and Structure Analysis of the V L and V H Genes from Anti hTNF α Hybridoma Cell Line

作　　者：陈建军[1] 徐皓[1] 高长寿[1] 孙苗[1] 陈常庆[1] 

机构地区：[1]中国科学院上海生物工程研究中心

出　　处：《细胞与分子免疫学杂志》1997年第3期8-13,共6页Chinese Journal of Cellular and Molecular Immunology

摘　　要：从两株具有高亲和力及中和活性的抗人肿瘤坏死因子－α（ｈＴＮＦ－α）的单抗杂交瘤细胞株１Ｃ３、３Ｆ６中分别提取总ＲＮＡ，通过ＲＴ－ＰＣＲ扩增并克隆了抗体的ＶＬ、ＶＨ基因。核苷酸序列分析表明，所克隆基因均为抗体的ＶＬ、ＶＨ基因，并分别属于Ｋａｐｐａ链Ⅳ亚组和重链Ⅲ（Ｄ）亚组。此外，两抗体可变区分子模型及与ｈＴＮＦ－α的结合模型的建立及分析，为进一步的基因工程抗体研究及抗原－抗体相互作用机理的研究奠定了基础。Mouse anti hTNF α monoclonal antibody 1C3 and 3F6 displayed a high affinity and specificity for hTNF α The variable light and heavy chain region(V L and V H) genes from the two hybridomas were cloned and amplified with RT PCR The nucleotide sequence analysis indicated that the cloned genes encoded the variable light and heavy chain domains from mouse anti hTNF α respectively and belonged to Kappa chain Ⅳ subgroup and heavy chainⅢ(D) subgroup respectively Moreover,the construction and analysis of molecular model of two McAb variable region and binding model of hTNF α laid the foundation of the further researches of engineering Ab and mechanism of Ag Ab interaction

关 键 词：HTNF-Α 肿瘤坯死因子α 单克隆抗体 可变区基因 

分 类 号：R392-33[医药卫生—免疫学]