원문정보
초록
영어
In this study, we will attempt to analyze the ER localization, glycosylation pattern and anti-tumor activity of KDEL-fused monoclonal antibody. Advantages of the baculovirus insect cell expression system for production of recombinant proteins include high capacity, flexibility, and glycosylation capability. In this study, this expression system was exploited to produce anti-cancer monoclonal antibody (mAb) CO17-1A, which recognizes the antigen GA733 that is highly expressed on the surface membrane of colorectal carcinoma cells. The heavy chain (HC) and light chain (LC) genes of KDEL-fused mAb CO17-1A were cloned under the control of P10 and PPH promoters in the pFastBacTM dual vector, respectively. Gene expression cassettes carrying the HC and LC genes were transposed into a bacmid in Escherichia coli (DH10Bac). The transposed bacmid was transfected to Sf9 insect cells to generate baculovirus expressing mAb CO17-1A. In future, we will study the expression and localization of KDEL-fused mAb CO17-A in baculovirus infected cells by PCR, western blot, enzyme linked immunosorbent assay (ELISA), confocal immunofluorescence, fluorescence-activated cell sorting analysis (FACs) and antibody dependent cell cytotoxicity (ADCC).