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应用ＲＴ－ＰＣＲ技术从分泌抗人黑色素瘤单克隆抗体的杂交瘤细胞ＨＢ８７６０中克隆了抗体轻、重链可变区基因，然后用（Ｇｌｙ４Ｓｅｒ）３连接肽基因将ＶＨ、ＶＬ连接成ＳｃＦｖ基因，并进行了序列测定．计算机分析表明ＶＨ，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．ＶＨ、ＶＬ、ｌｉｎｋｅｒ拼接正确．ＳｃＦｖ基因全长７２９ｂｐ，其中ＶＨ基因长３６０ｂｐ，编码１２０个氨基酸，ＶＬ基因长３２４ｂｐ，编码１０８个氨基酸．在噬菌粒表达载体ｐＣＡＮＴＡＢ５Ｅ中表达了可溶性的ＳｃＦｖ蛋白，表达产物经流式细胞仪检测可特异地与黑色素瘤细胞结合，不与肝癌、胃癌及良性黑痣细胞结合
The light and heavy chain variable region genes were cloned from the HB8760 cells secreting anti-human melanoma monoclonal antibodies by RT-PCR. Then, the VH and VL genes were linked to the ScFv gene by the (Gly4Ser) 3 linker peptide, And sequenced. Computer analysis showed that both VH and VL are in line with the characteristics of the mouse antibody variable region and are functionally rearranged antibody variable region genes. VH, VL, linker splicing correct. The full length of ScFv gene is 729bp, in which the VH gene is 360bp long and encodes 120 amino acids. The VL gene is 324bp long and encodes 108 amino acids. The soluble ScFv protein was expressed in the phagemid expression vector pCANTAB5E. The expression of the expressed protein was detected by flow cytometry. It could specifically bind to melanoma cells and not to liver cancer, gastric cancer and benign mole cells