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)
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Keyword |
pET 22b (+) vector, single chain fragment variable (scFv), lym-1 antibody, Raji cell lymphoma |
Full text Article |
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