Human catalytic antibody Se‐scFv‐B3 with high glutathione peroxidase activity

Rui Huo; Jingyan Wei; Junjie Xu; Shaowu Lv; Qingchuan Zheng; Fei Yan; Jiaming Su; Jia Fan; Jieshuai Li; Yujing Duan; Yang Yu; Fenghai Jin; Weiguo Sun; Yi Shi; Dengli Cong; Wei Li; Ganglin Yan; Guimin Luo

doi:10.1002/jmr.903

journal article Jun 23, 2008

Human catalytic antibody Se‐scFv‐B3 with high glutathione peroxidase activity

Rui Huo Jingyan Wei Junjie Xu

Shaowu Lv Qingchuan Zheng Fei Yan

Jiaming Su Jia Fan

Jieshuai Li Yujing Duan Yang Yu

Fenghai Jin Weiguo Sun

Yi Shi

Dengli Cong Wei Li

Ganglin Yan Guimin Luo

Journal of Molecular Recognition Vol. 21 No. 5 pp. 324-329 · Wiley

View at Publisher Save 10.1002/jmr.903

Abstract

AbstractIn order to generate catalytic antibodies with glutathione peroxidase (GPX) activity, we prepared GSH‐S‐2,4‐dinitrophenyl t‐butyl ester (GSH‐S‐DNPBu) as target antigen. Three clones (A11, B3, and D5) that bound specifically to the antigen were selected from the phage display antibody library (human synthetic VH + VL single‐chain Fv fragment (scFv) library). Analysis of PCR products using gel electrophoresis and sequencing showed that only clone B3 beared intact scFv‐encoding gene, which was cloned into the expression vector pPELB and expressed as soluble form (scFv‐B3) in Escherichia coli Rosetta. The scFv‐B3 was purified by Ni2+‐immobilized metal affinity chromatography (IMAC). The yield of purified proteins was about 2.0–3.0 mg of proteins from 1 L culture. After the active site serines of scFv‐B3 were converted into selenocysteines (Secs) with the chemical modification method, we obtained the human catalytic antibody (Se‐scFv‐B3) with GPX activity of 1288 U/µmol. Copyright © 2008 John Wiley & Sons, Ltd.

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References

35

[1]

10.1038/35035189

[2]

10.1016/0925-4439(95)00091-7

[3]

10.1021/ja963602k

[4]

10.1177/002580240204200103

[5]

10.1016/s0899-9007(00)00570-0

[6]

10.1016/s0021-9258(18)53812-6

[7]

10.1016/0006-2952(92)90245-e

[8]

10.1080/15604280214279

[9]

10.1161/01.cir.0000026820.87824.6a

[10]

10.1128/mcb.22.11.3565-3576.2002

[11]

10.1021/tx990156g

[12]

Jiang ZH "The progress in mechanism of selenoprotein biosynthesis" Acta Biochim. Biophys. Sin. (2002)

[13]

10.1016/j.bbrc.2004.06.110

[14]

10.1006/bbrc.1998.8545

[15]

10.1039/a808347i

[16]

10.1016/s0167-4838(00)00130-8

[17]

10.1006/bbrc.1994.1175

[18]

10.2174/0929867033457502

[19]

10.1016/0891-5849(94)90079-5

[20]

10.1016/0006-2952(84)90083-2

[21]

10.1042/bj3590369

[22]

Selenium: Biochemical Role as a Component of Glutathione Peroxidase

J. T. Rotruck, A. L. Pope, H. E. Ganther et al.

Science 10.1126/science.179.4073.588

[23]

10.1016/j.jviromet.2004.11.021

[24]

10.1042/bj0281977

[25]

10.1016/0891-5849(93)90028-s

[26]

10.1006/abbi.2001.2551

[27]

10.1016/s0003-9861(03)00006-7

[28]

10.1074/jbc.m403032200

[29]

Takahashi K "Selenium‐dependent glutathione peroxidase protein and activity: immunological investigations on cellular and plasma enzymes" Blood (1986) 10.1182/blood.v68.3.640.640

[30]

10.1016/s1383-5718(99)00204-1

[31]

10.1016/0304-4165(85)90182-5

[32]

Wei JY "Screening of scFvs against cTnI from phage display antibody library and their expression in <!?A3B2 ts=19?> E. coli <!?A3B2 ts=0?> Rosetta" Chem. Res. Chinese U. (2005)

[33]

10.1177/1087057106287901

[34]

10.1021/ja00197a065

[35]

10.1016/j.cbpc.2005.05.003

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Details

Published: Jun 23, 2008
Vol/Issue: 21(5)
Pages: 324-329
License: View

Authors

Department of Sports Medicine Shanghai Jiao Tong University Affiliated Sixth People's Hospital Shanghai China

Department of Biological Sciences, Purdue University, West Lafayette, IN 47907;

Wenzhou University

North University of China

Cite This Article

Rui Huo, Jingyan Wei, Junjie Xu, et al. (2008). Human catalytic antibody Se‐scFv‐B3 with high glutathione peroxidase activity. Journal of Molecular Recognition, 21(5), 324-329. https://doi.org/10.1002/jmr.903

Human catalytic antibody Se‐scFv‐B3 with high glutathione peroxidase activity

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