摘要
利用PCR方法从抗CD2 0 ScFv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CD2 0 嵌合抗体Fab’片段表达载体 pYZFcd2 0 ,用pYZFcd2 0转化大肠杆菌 16C9,在 16C9菌中分泌表达可溶性抗CD2 0 Fab’片段 ,经分离纯化获得具有CD2 0 抗原特异结合活性的Fab’片段 ,竞争性免疫荧光抑制实验表明 ,抗CD2 0 Fab’片段能竞争性抑制亲本鼠源性抗CD2 0 单克隆抗体HI4 7和Daudi细胞CD2 0
The gene of the variable region of the heavy chain(V H) and light chain(V L) were amplified by PCR method from the expression vector of anti human CD 20 single chain Fv antibody fragment(ScFv),and then the expression vector (pYZFcd20)of chimeric antibody Fab' fragment against CD 20 was constructed by cloning V H,V L into the expression vector pYZF. The Fab' was expressed in Ecoli. The Fab' was purified by Protein G agarose. The competitive inhibition of immunofluorescence showed that the Fab' inhibited binding of the monoclonal antibody HI 47 to CD 20 on the surface of Daudi cell.
出处
《高技术通讯》
EI
CAS
CSCD
2000年第4期9-12,共4页
Chinese High Technology Letters
基金
863计划资助项目!(863102090303)
关键词
单克隆抗体
抗体Fab'
大肠杆菌
Monoclonal antibody, Chimeric antibody Fab' fragment, CD_(20)
