Earticle

초록

영어

Insect cell expression system using baculovirus has several benefits from high capacity, flexibility, safety to humans and glycosylation capability. Thus, the baculovirus insect cell system has been widely used for production of recombinant protein. In this study, the insect cell expression system was designed to produce anti-cancer mAb CO17-1A, which recognizes the antigen GA733 highly expressed on the surface membrane of colorectal carcinoma cells. The heavy (HC) and light chain (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 E.coli (DH10Bac). The bacmid was transfected to Sf9 insect cells to generate the baculovirus expressing mAb CO17-1A. Western blot and immunoflouorescence analyses 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 (optimum virus concentration) ranging (0.2, 1 and 5). Expression of mAb CO17-1A in insect cells significantly increased at both 48 and 72hr after transfection with the MOI 1. HPLC chromatography 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 human colorectal cancer 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.

저자

• Eun-Yi Oh [ Department of Biological Science, Wonkwang University, Iksan ]
• Kyung-Jin Lee [ Department of Biological Science, Wonkwang University, Korean Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea ]
• Da-young Park [ Department of Biological Science, Wonkwang University, Iksan ]
• Yangkang So [ Department of Biological Science, Wonkwang University, Iksan ]
• Zhe Lu [ Department of Biological Science, Wonkwang University, Iksan ]
• Mira song [ Department of Biological Science, Wonkwang University, Iksan ]
• Mi-Hyun Ahn [ Department of Biological Science, Wonkwang University, Iksan ]
• Doo-Byoung Oh [ Korean Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea ]
• Youngkwan Kim [ Department of Biological Science, Wonkwang University, Iksan ]
• Kisung Ko [ Department of Biological Science, Wonkwang University, Korean Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614