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Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity  

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))
Publication Information
Nuclear Medicine and Molecular Imaging / v.40, no.4, 2006 , pp. 211-217 More about this Journal
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.
Keywords
pET 22b(+)vector; single chain fragment variable(scFv); lym-1 antibody; Raji cell lymphoma;
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