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目的 :抗CD3单克隆抗体 (WuT3)可变区基因克隆及序列分析。方法 :采用RT PCR技术 ,从WuT3杂交瘤细胞总RNA中扩增VH 、VL 片段 ,经酶切后通过链接反应构建重组克隆载体 ,测序鉴定。结果 :通过国际联机检索发现VH 、VL 基因与Ig同源 ,分别符合小鼠IgVH 、Vκ基因特征。VH 基因属于鼠重链VH 第IIB亚类 ,全长 36 3bp ,可编码 12 1个氨基酸 ;VL 基因属于鼠κ轻链第III亚类 ,全长 330bp ,可编码 110个氨基酸。结论 :成功获得WuT3单抗的重、轻链可变区基因。
Objective: Cloning and sequence analysis of variable region gene of anti-CD3 monoclonal antibody (WuT3). Methods: VH and VL fragments were amplified by RT PCR from total RNA of WuT3 hybridoma cells. After digestion, the recombinant cloning vector was constructed and identified by sequencing. Results: The VH and VL genes were found homologous to Ig through international online search, which were in line with the IgVH and Vκ gene characteristics of mice respectively. The VH gene belongs to murine heavy chain VH subgroup IIB, which is 36 3bp in length and encodes 12 1 amino acids. The VL gene belongs to murine light chain III subgroup with a total length of 330 bp and can encode 110 amino acids. Conclusion: The heavy and light chain variable region genes of WuT3 mAb were successfully obtained.