人抗HBV-前S2抗体轻链基因的克隆及序列分析  

Cloning and sequence analysis of light chain gene of human antibody against HBV pre-S2

作　　者：石小枫[1] 刘杞[1] 王志毅[1] 张定凤[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,400010

出　　处：《中华肝脏病杂志》2001年第S1期30-33,共4页Chinese Journal of Hepatology

摘　　要：目的获得人源性乙型肝炎病毒前Ｓ２抗体轻链基因。方法用人工合成的乙型肝炎病毒前Ｓ２短肽（Ｐｒｅ－Ｓ２，１２０—１４５），从已构建的人源性噬菌体抗体库中，得到了针对Ｐｒｅ－Ｓ２阳性克隆，经竞争抑制实验证明其特异性和活性，合成一对轻链引物，经ＰＣＲ扩增，亚克隆入质粒ＰＵＣ１８进行序列测定及分析、结果筛选到的阳性克隆具有乙型肝炎病毒前Ｓ２特异性亲和力。构建了ｐＵＣ１８－ Ｐｒｅ－Ｓ２ｋ。轻链重组质粒，序列分析其为人ｋ轻链，包括了完整的恒定区和可变区，大小为 ６４５ ｂｐ。结论利用抗原抗体特异性反应原理。Objective To obtain the gene of light chain of human antibody against HBV pre-S2. Methods A human antibody against HBV pre-S2 displayed on phage was obtained from the constructed human immunoglobulin phage displaying library through four rounds panning of "adherence-elution - amplification" with Pre- S2 (120 aa- 145 aa) short peptide synthesized and help phage. The positive clone of anti-pre-s2 was characterized by ELISA competitive inhibition assay. The plasmid from the positive clone was amplified by PCR...

关 键 词：肝炎表面抗原 乙型 人抗体轻链 基因克隆 序列分析 乙型肝炎表面抗原前S2短肽 

分 类 号：R512.62[医药卫生—内科学]