Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity.
- Author:
Jae Ho JUNG
1
;
Tae Hyun CHOI
;
Kang Sun WOO
;
Wee Sup CHUNG
;
Soo Gwan KIM
;
Gi Jeong CHEON
;
Chang Woon CHOI
;
Sang Moo LIM
Author Information
1. Lab. of Nuclear Medicine, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul, Korea. larry@kcch.re.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
pET 22b (+) vector;
single chain fragment variable (scFv);
lym-1 antibody;
Raji cell lymphoma
- MeSH:
Animals;
Gamma Cameras;
Immunoglobulin G;
Kidney;
Liver;
Lymphoma*;
Mice;
Mice, SCID;
Periplasm;
Radionuclide Imaging;
Spleen
- From:Nuclear Medicine and Molecular Imaging
2006;40(4):211-217
- CountryRepublic of Korea
- Language:Korean
-
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 x 10(9) M(-1), respectively, and those of scFv lym-1 were 53.7% and 1.46 x 10(9) 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.
