title: A neutralizing scFv antibody against infectious bursal disease virus screened by flow cytometry.
abstract: Infectious bursal disease (IBD) is considered a vital viral disease that threatens the poultry industry worldwide. In this study, a recombinant single chain variable fragment (scFv) antibody library derived from chickens immunized with VP2 protein of infectious bursal disease virus (IBDV) was constructed. The library was subjected to three rounds of screening by flow cytometry (FCM) against VP2/IBDV through a bacteria display technology, resulting in the enrichment of scFvs. Three scFv clones with different fluorescence intensity were obtained by colony pick up at random. The obtained scFv antibodies were expressed and purified. Relative affinity assay showed the three clones had different sensitivity to VP2, in accordance with fluorescence activity cell sorting analysis (FACS). The potential use of the isolated IBDV-specific scFv antibodies was demonstrated by the successful application of these antibodies in Western blotting and ELISA assay. What's more, in vitro neutralization measurement showed that one of the three isolated antibodies possessed the neutralization function against IBDV. This study provides new strategies for screening of antibody library, and scFv antibodies isolated in this study may be utilized as lead candidates for further development of diagnostic or therapeutic antibodies for detection and treatment of IBDV infection.
authors.affiliation: College of Life Sciences and Agro-forest, Qiqihar University, Qiqihar 161000, PR China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Life Sciences and Agro-forest, Qiqihar University, Qiqihar 161000, PR China. Electronic address: 2731848980@qq.com.
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Xie
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology*; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D018175:Birnaviridae Infections / Q000276:immunology; D036022:Capsid Proteins / Q000276:immunology; D002645:Chickens; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D015723:Gene Library; D007243:Infectious bursal disease virus / Q000276:immunology*; D009500:Neutralization Tests; D011201:Poultry Diseases; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D036022:Capsid Proteins; D057127:Single-Chain Antibodies
keywords: Anchored periplasm expression; Antibody library; Fluorescence-activated cell sorting; Infectious bursal disease virus; Neutralizing antibody