Construction of Antigenized Antibodies Expressing an Immunodominant Epitope AA32-45 of the Hepatitis C Virus Core Protein.
By using site-directed mutagenesis, we created a unique Xho I site in the CDR3 of the heavy-chain variable domain (V(H)). Two antibody molecules, one carrying one or and the other two repeats of an immunodominant epitope AA32-45 (GVYLLPRRGPRLGV) of the hepatitis C virus core protein in CDR3 of V(H) were engineered and designated Ig-E1, Ig-E2 respectively. We found that both antigenized antibodies lost the HBsAg-binding ability and the insertion of one repeat of GVYLLPRRGPRLGV epitope into the CDR3 of the V(H) domain did not appreciably affect the H chain to assemble with L chain to form a stable H(2)L(2) tetramer. Ig-E1 was able to be recognized by the polyclonal antibody against AA1-58 of the core protein as characterized by Western blot. However, Ig-E2 could not be assembled into H(2)L(2) tetramer.
Yu
Yong-Jun
YJ
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China.
Wu
Xiang-Fu
XF