Construction of Single-chain Antibody to Carcinoembryonic Antigen and Its High Expression in E. coli.
The heavy and light chain variable region (V(H) and V(K)) genes encoding the murine monoclonal antibody E(7)B(10) to carcinoembryonic antigen were linked into single-chain antibody gene with 36 oligonucleotide (Gly(3)Ser)(3) by using recombinant PCR. The construct was highly expressed in E. coli as inclusion bodies (IB), accounting for 20% of the total bacteria proteins, and was characterized by SDS-PAGE and Western blot. When the content of inclusion bodies was denatured and followed by renaturation, it was shown to possess abiliby to kind to its specific antigen CEA by using RIA and competitive ELISA.
Li
Biao
B
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. xfwu@sunm.shcnc.ac.cn
Zhu
Cheng-Mo
CM
Wu
Xiang-Fu
XF