Nucl Med Mol Imaging.  2006 Aug;40(4):211-217.

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

• Jung JH
• Choi TH
• Woo KS
• Chung WS
• Kim SG
• Cheon GJ
• Choi CW
• Lim SM

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.

Keyword

pET 22b (+) vector; single chain fragment variable (scFv); lym-1 antibody; Raji cell lymphoma

MeSH Terms

Animals
Gamma Cameras
Immunoglobulin G
Kidney
Liver
Lymphoma*
Mice
Mice, SCID
Periplasm
Radionuclide Imaging
Spleen
Immunoglobulin G