HBV Pre S2 3B9单抗轻、重链可变区基因的分离,序列分析及单链可变区抗体基因的构建  

AMPLIFICATION AND SEQUENCING OF THE V_H/V_L GENES OF 3B9 McAb WITH SPECIFICITY FOR HBV-PRE S2 AND CONSTRUCTION OF A ScFv ANTIBODY GENE

作　　者：程云[1] 周继文[1] 罗清华[1] 杨守纯[1] 韩风连 戎广亚[1] 张建宗[1] 乔小江 曹阳[1] 

机构地区：[1]解放军302医院免疫研究室,北京100039

出　　处：《中华微生物学和免疫学杂志》1995年第3期149-152,共4页Chinese Journal of Microbiology and Immunology

基　　金：全军85青年基金资助

摘　　要：3B9(杂交瘤细胞)是一株分泌HBV Pre S2抗原的单抗细胞,为从分子水平分析、了解3B9株进而为制备新一代基因工程抗体奠定基础,用分子生物学技术,提取3B9杂交瘤细胞总RNA,经反转录、PCR分离获得了3B9单抗的轻、重链可变区基因,并利用PCR及双脱氧核苷酸链末端终止法对3B9单抗的可变区基因的核苷酸序列进行了分析。结果表明,3B9单抗轻链隶属小鼠Ig Kappa轻链第Ⅱ亚组,JK_1基因重排;而3B9单抗重链隶属小鼠Ⅰg重链第Ⅱc亚组。在此基础上,作者用一编码疏水性多肽接头的DNA片断将3B9单抗轻、重链可变区基因连接,成功地构建了3B9单抗单链可变区抗体基因。为下一步克隆、表达有抗原结合活性的单链抗体打下了良好的物质基础。The rearranged Vnand VLgenes from 3B9 McAb were amplified and sequenced by PCR. The 351bp and 327bp of the 3B9 McAb VHand VL genes were analysed. These results showed that 3B9 VH/VI. genes belonged to themouse Ig heavy and light chain subgroup I c and I ,respectively. The VH and VL genes were ligated into a single chain Fv antibody (ScFv) gene by a specific linker encoding a flexible peptide. The restriction sites in the 3B9 ScFv are for the clone in the expressive phagemid vector.

关 键 词：序列分析 单链抗体基因 聚合酶链反应 

分 类 号：R392.11[医药卫生—免疫学]