Separate Expression and Characterization of the Heavy- and the Light-chain of A HBsAg Specific Murine-human Chimeric Antibody in Insect Cells.
Both the heavy- and light-chain of a murine-human chimeric antibody with specificity for hepatitis B virus surface antigen have been expressed separately in a baculovirus expression system. The heavy- and light-chain variable region (VH and VL) genes encoding the marine-monoclonal antibody OH3 were combined with human gamma3 and kappa constant region genes to construct the murine-human chimeric genes, respectively. The transfer vectors containing the chimeric genes were cotransfected into Sf9 cells with linearized virus DNA and recombinant viruses were isolated by dot hybridization, PCR and Southern blot analysis. The chimeric heavy- and light-chain was expressed respectively in the recombinant viruses-infected insect cells and the characterization by Western blot and competitive ELISA demonstrated that both of the expressed chimeric heavy- and light-chain have the specificity of HBsAg binding.
Yu
Yong-Jun
YJ
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China.
Chen
Ya-Hua
YH
Jiang
Yu-Lei
YL
Wu
Xiang-Fu
XF