抗人小细胞肺癌单抗4B3重、轻链可变区基因的体外扩增、克隆和序列分析

目的　获得抗人小细胞肺癌单抗 4B3的重 ,轻链可变区基因。方法　根据小鼠免疫球蛋白可变区骨架区的保守序列分别设计了 4B3单抗重 ,轻链的可变区基因体外扩增的两对引物 ,从体外培养的 4B3杂交瘤细胞中提取总RNA ,反转录成cD NA ,以cDNA为模板 ,分别用设计的两对引物进行PCR扩增 ,扩增出的重 ,轻链基因片段 ,分别插入载体 pT Adv中 ,筛选出阳性克隆 ,并测序、分析。结果　重链可变区基因全长 345bp ,编码 115个氨基酸 ,轻链可变区基因全长 315bp ,编码 10 5个氨基酸。结论　上述研究为 4B3单抗的进一步改造和利用提供了实验基础。

Amplification, cloning and sequence analyses of the VH and VL genes of an anti-Human small cell lung can-cer monoclonal antibody 4B3

Objective To obtain V H and V L genes of mAb 4B3 against human small cell lung cancer. Methods By comparing the conserved regions at murine germ-line genes encoding V regions of immunoglobulin heavy chain and light chain, Two pairs of primers were designed for the amplification of V H and V L genes respectively. Using RT-PCR, the genes of the heavy and light chain variable regions (V H and V L) of mAb 4B3 were obtained from anti-human small cell lung cancer hybridoma 4B3. Results The V H gene is 345bp and encodes 115 amino acids, the V L gene is 315bp and encodes 105 amino acids. Conclusion This study will supply a foundation for increasing reform and application of 4B3 monoclonal antibody.