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

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

In 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.

Original languageEnglish (US)
Pages (from-to)323-328
Number of pages6
JournalJournal of Molecular Recognition
Volume21
Issue number5
DOIs
StatePublished - Sep 1 2008
Externally publishedYes

Fingerprint

Catalytic Antibodies
Immunoglobulin Variable Region
Single-Chain Antibodies
Glutathione Peroxidase
Clone Cells
Selenocysteine
Antigens
Affinity Chromatography
Bacteriophages
Serine
Electrophoresis
Catalytic Domain
Esters
Proteins
Gels
Metals
Escherichia coli
Polymerase Chain Reaction
Antibodies

Keywords

  • Antibody library
  • Catalytic antibody
  • Chemical modification
  • Glutathione peroxidase (gpx)
  • Selenium
  • Single-chain fv fragment (scfv)

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Huo, R., Wei, J., Xu, J., Lv, S., Zheng, Q., Yan, F., ... Luo, G. (2008). Human catalytic antibody Se-scFv-B3 with high glutathione peroxidase activity. Journal of Molecular Recognition, 21(5), 323-328. https://doi.org/10.1002/jmr.903

Human catalytic antibody Se-scFv-B3 with high glutathione peroxidase activity. / Huo, Rui; Wei, Jingyan; Xu, Junjie; Lv, Shaowu; Zheng, Qingchuan; Yan, Fei; Su, Jiaming; Fan, Jia; Li, Jieshuai; Duan, Yujing; Yu, Yang; Jin, Fenghai; Sun, Weiguo; Shi, Yi; Cong, Dengli; Li, Wei; Yan, Ganglin; Luo, Guimin.

In: Journal of Molecular Recognition, Vol. 21, No. 5, 01.09.2008, p. 323-328.

Research output: Contribution to journalArticle

Huo, R, Wei, J, Xu, J, Lv, S, Zheng, Q, Yan, F, Su, J, Fan, J, Li, J, Duan, Y, Yu, Y, Jin, F, Sun, W, Shi, Y, Cong, D, Li, W, Yan, G & Luo, G 2008, 'Human catalytic antibody Se-scFv-B3 with high glutathione peroxidase activity', Journal of Molecular Recognition, vol. 21, no. 5, pp. 323-328. https://doi.org/10.1002/jmr.903
Huo, Rui ; Wei, Jingyan ; Xu, Junjie ; Lv, Shaowu ; Zheng, Qingchuan ; Yan, Fei ; Su, Jiaming ; Fan, Jia ; Li, Jieshuai ; Duan, Yujing ; Yu, Yang ; Jin, Fenghai ; Sun, Weiguo ; Shi, Yi ; Cong, Dengli ; Li, Wei ; Yan, Ganglin ; Luo, Guimin. / Human catalytic antibody Se-scFv-B3 with high glutathione peroxidase activity. In: Journal of Molecular Recognition. 2008 ; Vol. 21, No. 5. pp. 323-328.
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AU - Huo, Rui

AU - Wei, Jingyan

AU - Xu, Junjie

AU - Lv, Shaowu

AU - Zheng, Qingchuan

AU - Yan, Fei

AU - Su, Jiaming

AU - Fan, Jia

AU - Li, Jieshuai

AU - Duan, Yujing

AU - Yu, Yang

AU - Jin, Fenghai

AU - Sun, Weiguo

AU - Shi, Yi

AU - Cong, Dengli

AU - Li, Wei

AU - Yan, Ganglin

AU - Luo, Guimin

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AB - In 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.

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