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论文类型：期刊论文

发表时间：2016-06-01

发表刊物：INTERNATIONAL IMMUNOPHARMACOLOGY

收录刊物：SCIE、PubMed、Scopus

卷号：35

页面范围：149-154

ISSN号：1567-5769

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

摘要：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 SF(v)6 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. (C) 2016 Elsevier B.V. All rights reserved.