대한핵의학회지 (1967년~2009년)
Nucl Med Mol Imaging 2006;40(4)211~217
pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가
(Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity)
Author 정재호1,최태현1,우광선1,정위섭1,김수관2,천기정1,3,최창운1,3,임상무1,3,
Jae Ho Jung, M.S.1, Tae Hyun Choi, Ph.D.1, Kang Sun Woo1, Wee Sup Chung1,Soo Gwan Kim2, Gi Jeong Cheon, M.D.1,3, Chang Woon Choi, M.D.1,3, and Sang Moo Lim, M.D.1,3
Affiliation 원자력의학원 핵의학연구실1, 방사선의학연구센터2, 핵의학과3
1Lab. of Nuclear Medicine, 2Radiological and Medical Sciences Research Center, 3Dept. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul, Korea
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 from using pET vector system. Immunoreactivity and affinity constant of IgG lym-1 were 54% and 1.83¡¿109 M-1, respectively, and those of scFv lym-1 were 53.7% and 1.46¡¿109 M-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. These results suggest that scFv lym-1 antibody can be useful for tumor imaging agent. (Nucl Med Mol Imaging 2006;40(4):211-217)

Keyword pET 22b (+) vector, single chain fragment variable (scFv), lym-1 antibody, Raji cell lymphoma
Full text Article 4004211.pdf 4004211.pdf
(03121)서울시 종로구 지봉로 29 금호팔레스빌딩 1705호
TEL : 02-745-2040 FAX : 02-745-3833 E-mail : ksnm@ksnm.or.kr
Copyright 2012 by The Korean Society of Nuclear Medicine(KSNM)