한국가금학회지 (Korean Journal of Poultry Science)

  • 제38권4호
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  • Pages.323-330
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  • 2011
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  • 1225-6625(pISSN)
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  • 2287-5387(eISSN)
한국가금학회 (The Korean Society of Poultry Science)
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가금 콕시듐증을 일으키는 Eimeria spp.의 포자충 항원에 결합하는 닭의 재조합 항체(ScFv)의 개발

The Development of Chicken Recombinant Single-chain Fv (ScFv) Antibody Reactive with Sporozoite Antigen of Eimeria spp. which Causes Avian Coccidiosis

  • 박동운 (창원대학교 자연과학대학 미생물학과) ;
  • 김언동 (창원대학교 자연과학대학 미생물학과) ;
  • 김성헌 (창원대학교 자연과학대학 미생물학과) ;
  • 한재용 (서울대학교 농업생명과학대학 농생명공학부 WCU 바이오모듈레이션) ;
  • 김진규 (창원대학교 자연과학대학 미생물학과)
  • Park, Dong-Woon (Department of Microbiology, College of Natural Sciences, Changwon National University) ;
  • Kim, Eon-Dong (Department of Microbiology, College of Natural Sciences, Changwon National University) ;
  • Kim, Sung-Heon (Department of Microbiology, College of Natural Sciences, Changwon National University) ;
  • Han, Jae-Yong (WCU Biomodulation, Department of Agricultural Biotechnology, and Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Kim, Jin-Kyoo (Department of Microbiology, College of Natural Sciences, Changwon National University)
  • 투고 : 2011.10.04
  • 심사 : 2011.11.22
  • 발행 : 2011.12.31
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초록

닭의 단일 클론 항체인 13C8 항체는 조류의 콕시듐증을 유발하는 것으로 알려진 Eimeria acervulina의 포자충(sporozoites) 항원에 결합하는 닭 항체이다. 그러나 닭 하이브리도마 유전자의 불안정성 때문에 분비되는 항체의 생산량이 낮은 단점이 있다. 이러한 단점을 보완하기 위해 hybridoma로 부터 항체의 중사슬 가변 부위(VH)유전자와 경사슬 가변 부위(VL) 유전자를 증폭하여 linker peptide로 연결해준 재조합 ScFv 항체 유전자를 구축하고, E. coli를 형질 전환시켜 재조합 단백질로 발현 정제하였다. ELISA 분석 결과 재조합 13C8 ScFv 항체는 세 종류의 Eimeria spp.에 대해 모두 항원 결합력을 나타내었으며, 염기서열 분석을 수행하여 germline sequence와 비교한 결과 닭 항체유전자의 다양성(diversity)은 pseudogene들의 gene conversion 기작에 의해 이루어짐을 제시해 주었다.

The chicken monoclonal antibody (mAb), 13C8, reacts with sporozoite antigens of Eimeria spp. which causes avian coccidiosis. Since this mAb was produced at low amount due to genetic instability of chicken hybridoma, a recombinant 13C8 single-chain Fv (ScFv) antibody was constructed by amplification of the variable domain of heavy (VH) and light chain (VL) genes of antibody derived from chicken hybridoma. The constructed 13C8 ScFv was successfully expressed in E. coli and purified as a soluble form. In ELISA analysis, this recombinant 13C8 ScFv antibody showed antigen binding activity as the original mAb. In addition, nucleotide sequence comparison of 13C8 gene to the germline chicken VL and VH genes suggested that the gene conversion with $V{\lambda}$ and VH pseudogenes might contribute to the diversification of VL and VH genes in chickens.

키워드

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