Generation of selenium-containing human single-chain Fv antibody with GPX activity

被引：0

作者：

Huo Rui ^{[1
,2
]}

Shi Yi ^{[1
]}

Wei Jing-Yan ^{[1
,2
]}

Xu Jun-Jie ^{[1
]}

Lue Shao-Wu ^{[2
]}

Yan Fei ^{[2
]}

Su Jia-Ming ^{[3
]}

Duan Yu-Jing ^{[1
]}

Wang Shi-Wen ^{[1
]}

Cong Deng-Li ^{[1
]}

Li Wei ^{[3
]}

Yan Gang-Lin ^{[2
]}

Luo Gui-Min ^{[2
]}

机构：

[1] Jilin Univ, Coll Pharmaceut Sci, Changchun 130021, Peoples R China

[2] Jilin Univ, Key Lab Mol Enzymol & Engn, Minist Educ, Changchun 130021, Peoples R China

[3] Jilin Univ, Coll Life Sci, Changchun 130021, Peoples R China

来源：

CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE | 2008年 / 29卷 / 07期

关键词：

human scFv; phage display antibody library; GPX; selenium; chemical mutation;

D O I：

暂无

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Human single chain fragments variable(scFvs) against GSH were screened from a phage display human scFv antibody library. After three rounds of panning, five clones (2, 11, 16, 24, 32) binding to GSH were selected by ELISA. Analysis of PCR products using gel electrophoresis and sequencing showed that three clones( 11, 16, 24) contained intact scFv-encoding gene. The scFv-encoding gene from clone 11 was sub-cloned into the expression vector pPELB and expressed as soluble form(scFv-11) in Escherichia coli Rosetta. The scFv-11 was purified by Ni2+-immobilized metal affinity chromatography. The specificity to GSH of scFv-11 was verified by immuno-dot blot. Selenium-containing human scFv (Se-scFv-11) with GPX activity of 351 U/mol was obtained by the chemical modification of the reactive serine residues into Secs.

引用

页码：1379 / 1383

页数：5

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