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Expression of Anti-colorectal Cancer Monoclonal Antibody CO17-1A in Insect Cells.

첫 페이지 보기
  • 발행기관
    한국당과학회 바로가기
  • 간행물
    한국당과학회 학술대회 바로가기
  • 통권
    2008 Eastern Asian Glycoscience Symposium (2008.11)바로가기
  • 페이지
    pp.93-93
  • 저자
    Mi-Hyun Ahn, Mira Song, Kyung-Jin Lee, Eun-Yi Oh, Zhe Lu, Da-Young Park, Yang-Kang So, Doo-Byoung Oh, Kisung Ko
  • 언어
    영어(ENG)
  • URL
    https://www.earticle.net/Article/A192370

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원문정보

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, we designed the insect cell expression system 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 transfered into a parent bacmid in E.coli (DH10Bac). The bacmid was transfected to Sf9 insect cells to generate the baculovirus expressing mAb CO17-1A (CO17-1A-Bac virus). Western blot and immunoflouorescence confocal analyses confirmed the mAb CO17-1A expression in CO17-1A-Bac virus infected insect cells. The optimum conditions for mAb expression were validated at 24, 48 and 72h 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 72h after transfection with the MOI 1. HPLC chromatography revealed that the mAb CO17-1A expressed in the insect cell 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

  • Mi-Hyun Ahn [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Mira Song [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Kyung-Jin Lee [ Department of Biological Science, College of Natural Sciences, Wonkwang University, Korea Research Institute of Bioscience and Biotechnology ]
  • Eun-Yi Oh [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Zhe Lu [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Da-Young Park [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Yang-Kang So [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Doo-Byoung Oh [ Korea Research Institute of Bioscience and Biotechnology, Daejeon ]
  • Kisung Ko [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]

참고문헌

자료제공 : 네이버학술정보

간행물 정보

발행기관

  • 발행기관명
    한국당과학회 [Korean Society for Glycoscience]
  • 설립연도
    2006
  • 분야
    의약학>약학
  • 소개
    본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.

간행물

  • 간행물명
    한국당과학회 학술대회
  • 간기
    연간
  • 수록기간
    2006~2022
  • 십진분류
    KDC 517 DDC 614

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