论文部分内容阅读
利用PCR方法从抗CD20ScFv表达载体上扩增重链可变区(VH)、轻链可变区(VL)基因, 然后将VH , VL基因重组到Fab′表达载体中, 构建成抗CD20 嵌合抗体Fab′片段表达载体pYZFcd20. 用pYZFcd20转化大肠杆菌16C9, 在16C9菌中分泌表达可溶性抗CD20Fab′片段, 经分离纯化获得具有CD20抗原特异结合活性的嵌合抗CD20抗体Fab′片段. 竞争性免疫荧光抑制实验表明, 抗CD20Fab′片段能竞争性抑制亲本鼠源性抗CD20单克隆抗体HI47和Daudi细胞CD20结合. MTT法测定结果显示, 抗CD20 Fab′对Daudi细胞生长具有抑制作用.
The heavy chain variable region (VH) and light chain variable region (VL) genes were amplified from the anti-CD20ScFv expression vector by PCR, and then the VH and VL genes were recombined into the Fab’ expression vector to construct an anti-CD20 chimera. The antibody Fab′ fragment expression vector pYZFcd20 was transformed into E. coli 16C9 with pYZFcd20, and the soluble anti-CD20 Fab′ fragment was secreted and expressed in 16C9 bacteria. The chimeric anti-CD20 antibody Fab′ fragment with specific binding activity of CD20 antigen was isolated and purified. Fluorescence inhibition experiments showed that the anti-CD20 Fab′ fragment could competitively inhibit the binding of the parental mouse anti-CD20 monoclonal antibody HI47 and Daudi cell CD20. MTT assay results showed that anti-CD20 Fab′ had an inhibitory effect on the growth of Daudi cells.