多药耐药性肿瘤细胞相关噬菌体抗体库的构建

Construction of Phage Antibody Library Associated with Multidrug Resistance in Carcinoma Cells

利用噬菌体抗体库技术 ,构建了多药耐药性肿瘤细胞相关噬菌体抗体库。以多药耐药性肿瘤细胞 KB- A1为免疫原免疫 Bal B/c小鼠 ,从其脾脏提取总 RNA,并分离出 m RNA。RT- PCR扩增出 VH 和 VL 基因 ,经重叠延伸反应组装成单链抗体 ( Sc Fv)。将其克隆入噬菌粒载体p CANTAB 5 E中 ,电转化 E. coli TG1 ,成功构建了库容为 0 .8× 1 0 6的噬菌体抗体库 。

Using phage antibody library technique, a phage antibody library associated with multidrug resistance in carcinoma cells was constructed. Three BalB/c mice were immunized with multidrug resistant carcinoma cell line KB-A1. The mRNA was purified from total RNA of spleen. The VH and VL genes were amplified by RT-PCR, and ScFv was assembled by splicing overlap extension PCR. These fragments were cloned into the phagemid pCANTAB 5E. A phage antibody library containing 0.8×10 6 individual clones was constructed. It would be useful to isolate specfic ScFv against the cell surface molecules associated with multidrug resistance in carcinoma cells.