抗慢性淋巴细胞白血病独特型单抗SM6可变区基因序列分析

DNA sequencing of variable regions of anti-idiotypic monoclonal antibody SM6 to BCLL

目的：对独特型抗体可变区基因进行序列分析，为其基因工程抗体的构建打下基础。方法：采用ＴＲＩｚｏｌＲ试剂从分泌抗独特型抗体ＳＭ６杂交瘤细胞株中提取ｍＲＮＡ，以特异性引物采用ＰＣＲ技术扩增并克隆出独特型单抗ＳＭ６重、轻链可变区（ＶＨ、ＶＬ）基因。将ＶＨ和ＶＬ基因分别克隆到ｐＧＥＭＴ载体中。采用链末端终止法荧光染色测定ＶＨ和ＶＬ基因序列，应用ＤＮＡＳＩＳ７分析软件并与美国国立卫生研究院基因库比较分析。结果：５个克隆的重链基因序列一致，３个克隆的轻链基因中有２个克隆基因一致。结论：ＶＨ和ＶＬ基因分别属Ｑ５２、ＪＨ３和ＶＫ１９、ＪＫ２。

Objective:To analyze the variable region sequences of an antiidiotypic monoclonal antibody to human chronic B cell leukemia (BCLL). Methods:The variable region sequences of light and heavy chains (VL、VH) of antiidiotypic McAb SM6 to BCLL were analyzed by their cDNA cloning and sequencing. Total RNA was extracted from a hybridoma cell line producing SM6. The VH and VL genes were amplified by RTPCR with specific primers. The PCR products were cloned into pGEMT vectors, then transfected into JM109. Results and Conclusion: It was confirmed by DNASIS7 sequence analysis that the fulllength and potentially functional genes were successfully cloned. The VH gene expressed in SM6 belongs to Q52, the JH region using the JH3. The VL gene belongs to VK19, the JK region using JK2.