抗CD3单克隆抗体重、轻链可变区基因的克隆及序列分析  

Cloning and sequence analysis of V_H and V_L gene of anti-CD3 McAb

作　　者：张囡[1] 陈明洁[1] 陈敬[1] 金海陵[1] 张爱华[1] 闭兰[1] 左建民[1] 王志友[1] 史良如[1] 

机构地区：[1]武汉生物制品研究所免疫研究室,武汉430060

出　　处：《中国免疫学杂志》2002年第9期608-610,共3页Chinese Journal of Immunology

摘　　要：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:To amplify and sequence of the variable region genes of anti CD3 McAb.Methods:The V H?V L genes were amplified by RT PCR from total RNA that were extracted from WuT3 hybridoma.Recombinant cloning vector was constructed and sequenced after the enzyme digestion.Results:It showed that V H gene consisted of 363 bp encoding amine acid residues,belongs to mouse heavy chain subgroup IIB;V L gene consisted of 330 bp encoding amine acid residues,belongs to mouse κ light chain subgroup III.Comparing with Kabat database,the V H?V L genes were in agreement with the characterization of DNA sequences present in the mouse Ig V H?V L regions respectively.Conclusion:The success of cloning of the V H?V L genes of WuT3 McAb lay a good foundation for the construction and expression of chimeric antibody.

关 键 词：McAb WuT3 基因克隆 DNA序列分析 抗CD3单克隆抗体 

分 类 号：R392－33[医药卫生—免疫学]