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应用ＲＴＰＣＲ技术，从分泌抗人肝癌单克隆抗体（ｍＡｂ）的杂交瘤细胞ＨＡｂ１８中，扩增出抗体ＶＨ和ＶＬ基因，并测序。计算机分析表明，ＶＨ和ＶＬ均符合小鼠抗体可变区的特征，为功能性重排的抗体可变区基因。用（Ｇｌｙ４Ｓｅｒ）３连接肽基因，将ＶＨ和ＶＬ连接成ＳｃＦｖ基因，并进行序列测定，结果表明，ＶＨ，ＶＬ，ｌｉｎｋｅｒ拼接正确，基因全长为７２６ｂｐ。用噬菌粒表达载体ｐＣＡＮＴＡＢ５Ｅ在大肠杆菌中表达了可溶性的ＳｃＦｖ融合蛋白，经流式细胞仪分析表达产物可特异地与肝癌细胞结合，而不与正常肝细胞及胃癌细胞结合
The VH and VL genes of antibody were amplified by RT-PCR from the HAb18 cells secreting anti-human hepatoma monoclonal antibody (mAb) and sequenced. Computer analysis shows that both VH and VL are in line with the characteristics of mouse antibody variable regions, and are functional rearranged antibody variable region genes. The peptide gene was ligated with (Gly4Ser) 3, the VH and VL were ligated into ScFv gene, and the sequence was determined. The results showed that the VH, VL and linker were spliced correctly, and the full-length gene was 726bp. The phagemid expression vector pCANTAB5E was used to express soluble ScFv fusion protein in E.coli. The expression product could be specifically bound to hepatoma cells by flow cytometry but not to normal hepatocytes and gastric cancer cells