Fusion and Expression of the Genes Encoding Murine-Human Chimeric Heavy Chain to Carcinoembryonic Antigen with Core-streptavidin Gene.
In order to reduce the human anti-murine antibody (HAMA) in radioimmunotherapy, the gene encoding the heavy chain variable region (V(H)) of the murine monoclonal antibody with high specificity and affinity to carcinoembryonic antigen was fused to the human C(gamma3) gene to construct the murine-human chimeric heavy chain antibody gene, then was linked to core-streptavidin which can specifically bind to biotin, facilitating its purification and radioisotope labeling. The fusion gene was expressed in E.coli at high level, accounting for 24% of the total bacteria protein, and was characterized by SDS-PAGE and Western-blots. When using RIA the content of inclusion bodies was denatured and followed by renaturation, it was shown by using RIA to possess ability to bind to its specific antigen of CEA. Using horseradish peroxidase (HRP) labeled biotin as antibody in Western-blots, one band of 70 kD only was detected, demonstrating the fusion protein not only had the ability to bind CEA, but also could bind biotin specifically.
Yang
Wei-Dong
WD
Shanghai Institute of Biochemistry, the Chinese Academy of Science, Shanghai 200031, China. xfwu@sunm.shcnc.ac.cn
Li
Biao
B
Zhu
Cheng-Mo
CM
Liao
Wen-Tao
WT
Yang
Guan-Zhen
GZ
Wu
Xiang-Fu
XF