新的人抗HBsAg抗体Fab cDNA的克隆及序列分析

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），并进行基础序列分析。方法 对噬菌体抗全库进行三轮“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的抗休天经地义人得到的抗惭肝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 analys is of human anti-HBs antigen Fab gene

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 sequence 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, competition inhibition ELISA and DNA sequencing. CONCLUSION Specific phage antibodies (Fab fragments) may lay a solid foundation for further research into their expression and purification.