抗血管内皮生长因子165抗体轻重链可变区基因的克隆与序列分析

Cloning and Sequencing of the VL and VH Genes from the Anti-vascular Endothelial Growth Factor Mc Ab Hybridoma CellLine

目的 获得抗血管内皮生长因子 1 6 5 ( VEGF1 65)抗体的轻重链可变区基因。 方法 从分泌具有中和抗血管内皮生长因子 1 6 5活性的单抗杂交瘤细胞株Vm D1 1中提取总 RNA,经 RT- PCR扩增并克隆该单抗的轻、重链可变区基因并进行序列分析。结果 抗体的轻链可变区基因 32 1 bp,属于小鼠第 亚群 ;重链可变区基因 35 4bp,属于小鼠第 ( B)亚群。结论 所克隆的基因经核苷酸序列分析分别为抗体的轻、重链可变区基因。

Objective The light chain variable region ( VL ) and heavy chain variable region ( VH) genes from the anti- VEGF1 65 Mc Ab hybridoma cell line were cloned for construction of it′s chimeric antibody.Methods The VL and VH genes were amplified from the total RNA of the Vm D1 1 hybridoma with RT- PCR and cloned into vectors,and then analyzed by sequenc- ing.Results The32 0 bp VL gene belong to mouse kappa light chain subgroup V,the35 4 bp VH gene belong to mouse heavy chains subgroup ( B) .Conclusions The nucleotide sequence analysis indicated that the cloned genes coded the light and heavy chain variable re- gion of mouse respectively.