Soluble Expression, Purification and Characterization of Single-chain Fv Catalytic Antibody(sFv-2F3)

Chemical Research in Chinese Universities ›› 2004, Vol. 20 ›› Issue (3): 317-322.

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Soluble Expression, Purification and Characterization of Single-chain Fv Catalytic Antibody(sFv-2F3)

SUN Ye¹, LI Wei-jia¹, MA Ji-sheng^1,2, MU Ying¹, DU Xiu-bo¹, YAN Gang-lin¹, LUO Gui-min¹

1. Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130023, P. R. China;
2. Jilin Institate of Natural Medicine, Changchun 130012, P. R. China

Received:2001-01-01 Online:2004-06-24 Published:2011-08-06
Supported by:
Supported by the Foundation of the Ministry of Science and Technology of China("863" Project No.2001AA215315 and "973" Project, No.G2000078102) and the National Natural Science Foundation of China(No.202720106, 30200050) and Jilin University.

Abstract

Abstract: To find optimal conditions for expressing the soluble form of sFv-2F3 and to study the purification and property of its derivative Se-sFv-2F3, the preferred expression conditions were investigated by means of orthogonal design. These culture conditions included incubation temperature, inducer concentration, induction time and cell concentration. The evaluation of expression was accomplished by the analysis of whole cell lysates and the yield of soluble sFv-2F3 was calculated according to the analysis of Profinder(FTI-500,Pharmacia). The purification procedure was carried out via a two-step purification procedure consisting of ion-exchange chromatography, followed by immobilized metal affinity chromatography(IMAC). The antioxidant efficacy of Se-sFv-2F3 was demonstrated by the determination of the content of the main product of lipid peroxidation, MDA, the viability of cells and the activity of LDH. We obtained the preferred culture conditions to grow the engineered bacteria and the procedure for preparing soluble sFv-2F3 and confirmed the antioxidant efficacy of Se-sFv-2F3.

Key words: Se-sFv-2F3, Soluble expression, Glutathione peroxidase, Antioxidant

SUN Ye, LI Wei-jia, MA Ji-sheng, MU Ying, DU Xiu-bo, YAN Gang-lin, LUO Gui-min. Soluble Expression, Purification and Characterization of Single-chain Fv Catalytic Antibody(sFv-2F3)[J]. Chemical Research in Chinese Universities, 2004, 20(3): 317-322.

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Soluble Expression, Purification and Characterization of Single-chain Fv Catalytic Antibody(sFv-2F3)

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