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目的克隆ＨＡｂ２５肝癌单克隆抗体（ＭＣＡｂ）的可变区基因。方法ＲＴ－ＰＣＲ＄从分泌ＨＡｂ２５单克隆抗体的杂交瘤细胞中，扩增出ＶＨ和ＶＬ基因，双脱氧链终止法测定其核苷酸序列。结果ＶＨ和ＶＬ基因的两端均含有完整的引物序列，ＶＨ基因全长３６０ｈＰ，编码１２０个氨基酸，ＶＬ基因全长３３０ｈＰ，编码１１０个氨基酸，重、轻链可变区内均仅含单一开放读框，具有明显的抗体可变区特征，与基因数据库（ＧｅｎＢａｎｋ冲的序列进行比较分析，基因库中无相同的基因，ＶＨ基因与小鼠重链ＶＨ１８６．２家族同源性最高（８４．００％），属小鼠屹重链可变区９个家族中的第三族，ＶＬ基因与小鼠ｋａｐｐａ轻链ＭＭＩｇＧＫＡＶＡＧ基因同源性最高（８２．００％），属小鼠Ｉｇｋａｐｐｐａ轻链第ＩＶ组。结论克隆的ＨＡｂ２５ＭｃＡｂ的可变区基因为功能性重排的小鼠可变区基因。
Objective To clone the variable region gene of HAb25 liver cancer monoclonal antibody (MCAb). Methods RT-PCR$ VH and VL genes were amplified from hybridoma cells that secrete monoclonal antibodies against HAb25, and their nucleotide sequences were determined by dideoxy chain termination method. Results The VH and VL genes contained intact primer sequences at both ends. The full-length VH gene was 360hP, encoding 120 amino acids. The VL gene was 330hP in length and encoded 110 amino acids. The heavy and light chain variable regions contained only a single open sequence. The reading frame has obvious characteristics of antibody variable region. Compared with the gene database (GenBank), there is no identical gene in the gene library. The VH gene has the highest homology with the mouse heavy chain VH186.2 family (84. 00%) is the third group in the 9 families of the mouse variable heavy chain variable region. The VL gene has the highest homology with the mouse kappa light chain MMIgGKAVAG gene (82.00%) and belongs to the mouse Igkapppa light chain. Group IV. Conclusions The variable region gene of the cloned HAb25 McAb is a functional rearranged mouse variable region gene.