Characterization of N-glycan structures and biofunction of anti-colorectal cancer monoclonal antibody CO17-1A produced in baculovirus-insect cell expression system

摘要: Advantages of the baculovirus insect cell expression system for production recombinant proteins include high capacity, flexibility, and glycosylation capability. In this study, was exploited to produce anti-cancer monoclonal antibody (mAb) CO17-1A, which recognizes antigen GA733. The heavy chain (HC) light (LC) genes mAb CO17-1A were cloned under control P(10) Polyhedrin promoters in pFastBac dual vector, respectively. Gene cassettes carrying HC LC transposed into a bacmid Escherichia coli (DH10Bac). transfected Sf9 cells generate expressing CO17-1A. Confocal immunofluorescence Western blot analyses confirmed baculovirus-infected cells. optimum conditions evaluated at 24, 48, 72 h after virus infection an multiplicity 1. Expression significantly increased infection. HPLC analysis status revealed that insect-derived (mAb(I)) had specific glycan structures. ELISA showed purified mAb(I) from culture supernatant specifically bound SW948 human colorectal cancer Fluorescence-activated sorting that, although structures differed their mammalian counterparts, similarly interacted with CD64 (FcgammaRI) Fc IgG, compared interactions mammalian-derived mAb. These results suggest is able express, assemble, secrete biofunctional full size