Generation of Selenium-containing Human Single-chain Fv Antibody with GPX Activity

Abstract

Abstract: 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 subcloned 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.

Key words: Human scFv, Phage display antibody library, GPX, Selenium, Chemical mutation

CLC Number:

Q511

TrendMD:

HUO Rui^1,2, SHI Yi¹, WEI Jing-Yan^1,2*, XU Jun-Jie¹, Ldoi: .

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Generation of Selenium-containing Human Single-chain Fv Antibody with GPX Activity

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