Nuclear Medicine and Molecular Imaging

The Korean Society of Nuclear Medicine (대한핵의학회)
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Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity

pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가

  • Jung, Jae-Ho (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Choi, Tae-Hyun (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Woo, Kang-Sun (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Chung, Wee-Sup (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Kim, Soo-Gwan (Radiological and Medical Sciences Research Center, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Cheon, Gi-Jeong (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Choi, Chang-Woon (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS)) ;
  • Lim, Sang-Moo (Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences(KIRAMS))
  • 정재호 (원자력의학원 핵의학연구실) ;
  • 최태현 (원자력의학원 핵의학연구실) ;
  • 우광선 (원자력의학원 핵의학연구실) ;
  • 정위섭 (원자력의학원 핵의학연구실) ;
  • 김수관 (원자력의학원 방사선의학연구센터) ;
  • 천기정 (원자력의학원 핵의학연구실) ;
  • 최창운 (원자력의학원 핵의학연구실) ;
  • 임상무 (원자력의학원 핵의학연구실)
  • Published : 2006.08.31
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Abstract

Purpose: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. Methods: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. Results: An active scFv lym-1 could be produced in E. coli with soluble iron using PET vector system. Immuuoreaetivity and affinity constant of IgG lym-1 were 54% and $1.83{\times}10^9M^{-1}$, respectively, and those of scFv lym-1 were 53.7% and $1.46{\times}10^9M^{-1}$, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. Conclusions: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1 There results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

재조합된 scFv lym-1의 세포결합 실험에서 높은 면역반응성을 보였으며, 비특이 반응에서 모항체인 IgG lym-1에 의해 결합이 억제됨을 확인하였고, 동물 영상을 통해 IgG보다 분자랑이 작은 scFv lym-1 항체가 빠른 체내대사율과 종양섭취율을 보여 방사면역진단에 유용할 것으로 보여진다.

Keywords

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