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Construction and characterization of a highly reactive chicken-derived single-chain variable fragment (scFv) antibody against Staphylococcus aureus developed with the T7 phage display system.

Posted by on in 2016
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Li J1Xu Y2Wang X1Li Y1Wang L2Li X32016. Int Immunopharmacol. 35:149-54. doi: 10.1016/j.intimp.2016.02.024. Epub 2016 Apr 16.

1
School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China.
2
School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China; Ministry of Education Center for Food Safety of Animal Origin, Dalian 116620, China.
3
School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China; Ministry of Education Center for Food Safety of Animal Origin, Dalian 116620, China. Electronic address: xiaoyuli@dlut.edu.cn.

 

Abstract

The purpose of this study was to construct a single-chain variable fragment (scFv) antibody from chicken egg yolk immunoglobulin (IgY) by means of genetic engineering and subsequent panning for a specific antibody against Staphylococcus aureus.

METHODS AND RESULTS:

We amplified the scFv using blood and spleen obtained from 100-day-old Roman chickens immunized with inactivated S. aureus and subsequently constructed a T7 phage display antibody library using phage display technology. Four non-repeated blood scFv and 6 spleen scFv were obtained following 3 rounds of panning of the T7 phage display antibody library, enzyme-linked immunosorbent assay and sequencing. These 10 scFv were cloned into the prokaryotic expression vector pCold I with expression induced at a low temperature. Four soluble proteins were obtained. Among them, soluble protein SFV6 derived from the spleen showed good reactivity against S. aureus using indirect ELISA and produced a particularly strong antibacterial effect in vitro.

CONCLUSION:

We were successful in isolating a highly specific scFv antibody against S. aureus from the spleen phage display library. This study provides a simple and rapid method for the quick preparation of a large number of antibodies against S. aureus and provides the foundation for the positioning of antibodies in the organism and the study of the antibacterial mechanism through which the antibody functions.

KEYWORDS:

Chicken IgY; Phage display; Single-chain variable fragment (scFv); Staphylococcus aureus

PMID:
 
27046516
 
DOI:
 
10.1016/j.intimp.2016.02.024
[Indexed for MEDLINE]
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