关键词: 噬菌体抗体;肝炎表面抗原，乙型;人Ig可变区基因;序列分析

    摘 要:目的 从已构建的人抗-HBsAg噬菌体抗体库中筛选出结合乙肝表面抗原(HBsAg)的特异的人噬菌体抗体(Fab段)，并进行基因序列分析. 方法 对噬菌体抗体库进行三轮“吸附-洗脱-扩增”的筛选，使特异性噬菌体抗体得到了富集.ELISA实验和ELISA竞争抑制实验鉴定出乙肝表面抗原的特异性抗体，同时对筛选出的克隆进行基因序列分析. 结果 得到与乙肝表面抗原特异结合的抗体，并通过基因序列分析证明为抗乙肝HBsAg的抗体. 结论 得到的抗乙肝HB-sAg的特异抗体为下一步表达纯化工作奠定了基础.

    Screening of phage antibodies to hepatitis B surface antigen and analysis of human anti┐HBs antigen Fab gene

    LIAO Qi-Hua GAO Hui CHEN Nan-Chun YANG An-Gang CHEN Su-Min SU Kuan-Yuan YU Zhou-Yao

    1 Department of Biochemistry and Molecular Biolo-gy，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China，

    2 Hepatology Center of Chinese PLA，Air Force Guangzhou Hospital，Guangzhou510600，China

    Keywords:phage antibodies;hepatitis B surface antigens;human Ig verified region gene;sequence analysis Abstract:AIM To screen phage antibodies(Fab fragments)to hepatitis B surface antigen(HBsAg)from the constructed phage display library，and to analyse it with DNA sequencing and restriction enzyme mapping.METHODS Three rounds selection against coated HBsAg showed specific enrichment of phage antibodies.The specificity of the clones was verified by ELISA and competition inhibition ELISA.The nucleotide se-quence of the target DNA was determined by the dideoxy chain termination method.RESULTS Phage antibodies(Fab fragments)to hepatitis B surface antigen(HBsAg)were determined by ELISA ......