원문정보
초록
영어
Insect cell expression system using baculovirus has several benefits from high capacity, flexibility, and safety to humans and glycosylation capability. Thus, the baculovirus insect cell system has been widely used for production of recombinant protein. The HC and LC genes of the mAb CO17-1A were cloned under the control of two different promoters, P10 and PPH, respectively, on baculovirus expression pFastBacTM Dual vector. The gene expression cassettes carrying HC and LC genes were transferred into a parent bacmid in DH10Bac. Immunoflouorescence analyses and Western blot confirmed expression and secretion of mAb CO17-1A in the virus infected insect cells. The optimum condition for mAb expression was optimized at 24, 48 and 72hr after the virus infection with MOI ranging (0.2, 1 and 5). HPLC analysis revealed that the insect-derived mAb CO17-1A had insect specific glycan structures. Cell ELISA showed that the purified mAb from insect cell cultured media had a specific binding activity to SW948 cell. These results indicated that the baculovirus insect cell system is able to express, assemble, and secrete functional full size monoclonal antibody with insect specific glycosylation.