纤维蛋白单抗SZ-58嵌合Fab片段在大肠杆菌中的表达  

Construction and expression of chimeric Fab fragment of anti-fibrin monoclonal antibody SZ-58 in E .coli

作　　者：刘跃[1] 夏利军[1] 顾建明[1] 傅建新[1] 万海英[1] 阮长耿[1] 

机构地区：[1]江苏省血液研究所,苏州医学院血栓与止血研究室215007

出　　处：《中华微生物学和免疫学杂志》1996年第2期100-103,共4页Chinese Journal of Microbiology and Immunology

基　　金：江苏省卫生厅基金

摘　　要：为减少鼠源单抗的免疫原性，降低其分子量，应用基因重组技术将纤维蛋白单抗ＳＺ－５８可变区基因与人ＩｇＧ１恒定区基因Ｃ_Ｈ１、Ｃｋ进行拼接、扩增。将扩增后的ＳＺ－５８嵌合Ｆａｂ基因克隆至噬菌体质粒，并导入大肠杆菌中进行表达。表达的嵌合Ｆａｂ片段为可溶性。放免检测其在表达上清中的含量约为２００μｇ／Ｌ，免疫印迹电泳证实ＳＺ－５８嵌合Ｆａｂ片段能特异地与纤维蛋白Ｄ－Ｄｉｍｅｒ结合。In a previous study , an antifibrin monoclonal antibody (McAb ) SZ-58 was generated from mouse origin. Experiments showed in vitro that it had good fibrin clot binding characteristic. In order to reduce the immunogenicity of McAb and lower its molecular weight , the variable region gene of SZ-58 were fused to human IgGl - constant region gene CH1 , Ck. The constructor was cloned into phagemid vector pHEN-1-Fab/Hu for expressing the soluble form of chimeric SZ-58 Fab fragment in non-suppressor E. coli HB2151. The concentration of SZ-58 Fab fragment in expression supernatant was about 200μg/L measured by IRMA. Western blot showed that it had higher fibrin D-Dimer binding activity than its parent McAb.

关 键 词：单克隆抗体 纤维蛋白 嵌合Fab片段 大肠杆菌 

分 类 号：R392－33[医药卫生—免疫学]