抗人整合素ανβ_3 单抗E10鼠/人嵌合Fab噬菌体抗体的构建和表达  

Construction and expression of mouse/human chimeric Fab of mAb E10 against integrin ανβ_3 on phage

作　　者：王臣[1] 侯利华[2] 张彦明[1] 李建民[2] 廖振林[2] 杜桂鑫[2] 陈薇[2] 童贻刚[2] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《西北农林科技大学学报（自然科学版）》2004年第5期9-13,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家863计划项目资助(2001AA5361)

摘　　要：　为构建抗人整合素ανβ3单抗E10鼠/人嵌合Fab噬菌体抗体表达载体,并在噬菌体表面表达,采用PCR方法从整合素ανβ3单抗E10ScFv载体中,扩增重链可变区VH和轻链可变区VL基因。将VH基因与人重链恒定区CH1基因连接,VL基因与人CK基因连接,分别构建了鼠/人嵌合重链FD基因和轻链基因,将其克隆入pComb3,构建了鼠/人嵌合Fab噬菌体抗体表达载体,用辅助噬菌体VCSM13超感染,间接ELISA及竞争抑制ELISA检测鼠/人嵌合Fab噬菌体抗体活性。结果成功构建了鼠/人嵌合Fab噬菌体抗体表达载体,并在噬菌体表面展示了可结合人整合素ανβ3抗原的鼠/人嵌合Fab抗体。To construct and express the mouse/human chimeric Fab antibodies on phage,the genes of the variable region of the heavy chain (V_H) and light chain (V_L) were amplifyied by PCR from expression vector of anti-human integrin ανβ_3 ScFv,and then V_H and V_L genes of mAb E10 were ligated with human heavy chain C_H1 and human C_K genes.The chimeric Fab expression vector was constructed by cloning the chimeric heavy chain FD and light chain genes into pComb3.Fab antibodies were expressed on phage by superinfection with the helper phage VCSM13,and the binding activity of antibody was detected by indirect ELISA and competitive inhibition ELISA.Mouse/human chimeric Fab expression vector was constructed and expressed on phage successfully.The Fab antibodies had binding activity with human integrin ανβ_3 .

关 键 词：整合素ΑΝΒ3 噬菌体展示 PCR 肿瘤血管形成 细胞粘附分子 鼠/人嵌合Fab抗体 

分 类 号：R730[医药卫生—肿瘤] R392.11[医药卫生—临床医学]