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Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display

Fusarium verticillioides is the primary causal agent of Fusarium ear and kernel rot in maize, producing fumonisin mycotoxins that are toxic to humans and domestic animals. Rapid detection and monitoring of fumonisin-producing fungi are pivotally important for the prevention of mycotoxins from enteri...

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Autores principales: Hu, Zu-Quan, Liu, Jin-Long, Li, He-Ping, Xing, Shu, Xue, Sheng, Zhang, Jing-Bo, Wang, Jian-Hua, Nölke, Greta, Liao, Yu-Cai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3397510/
https://www.ncbi.nlm.nih.gov/pubmed/22837678
http://dx.doi.org/10.3390/ijms13067038
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author Hu, Zu-Quan
Liu, Jin-Long
Li, He-Ping
Xing, Shu
Xue, Sheng
Zhang, Jing-Bo
Wang, Jian-Hua
Nölke, Greta
Liao, Yu-Cai
author_facet Hu, Zu-Quan
Liu, Jin-Long
Li, He-Ping
Xing, Shu
Xue, Sheng
Zhang, Jing-Bo
Wang, Jian-Hua
Nölke, Greta
Liao, Yu-Cai
author_sort Hu, Zu-Quan
collection PubMed
description Fusarium verticillioides is the primary causal agent of Fusarium ear and kernel rot in maize, producing fumonisin mycotoxins that are toxic to humans and domestic animals. Rapid detection and monitoring of fumonisin-producing fungi are pivotally important for the prevention of mycotoxins from entering into food/feed products. Chicken-derived single-chain variable fragments (scFvs) against cell wall-bound proteins from F. verticillioides were isolated from an immunocompetent phage display library. Comparative phage enzyme-linked immunosorbant assays (ELISAs) and sequencing analyses identified four different scFv antibodies with high sensitivity. Soluble antibody ELISAs identified two highly sensitive scFv antibodies, FvCA3 and FvCA4, with the latter being slightly more sensitive. Three-dimensional modeling revealed that the FvCA4 may hold a better overall structure with CDRH3, CDRL1 and CDRL3 centered in the core region of antibody surface compared with that of other scFvs. Immunofluorescence labeling revealed that the binding of FvCA4 antibody was localized to the cell walls of conidiospores and hyphae of F. verticillioides, confirming the specificity of this antibody for a surface target. This scFv antibody was able to detect the fungal mycelium as low as 10(−2) μg/mL and contaminating mycelium at a quantity of 10(−2) mg/g maize. This is the first report that scFv antibodies derived from phage display have a wide application for rapid and accurate detection and monitoring of fumonisin-producing pathogens in agricultural samples.
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spelling pubmed-33975102012-07-26 Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display Hu, Zu-Quan Liu, Jin-Long Li, He-Ping Xing, Shu Xue, Sheng Zhang, Jing-Bo Wang, Jian-Hua Nölke, Greta Liao, Yu-Cai Int J Mol Sci Article Fusarium verticillioides is the primary causal agent of Fusarium ear and kernel rot in maize, producing fumonisin mycotoxins that are toxic to humans and domestic animals. Rapid detection and monitoring of fumonisin-producing fungi are pivotally important for the prevention of mycotoxins from entering into food/feed products. Chicken-derived single-chain variable fragments (scFvs) against cell wall-bound proteins from F. verticillioides were isolated from an immunocompetent phage display library. Comparative phage enzyme-linked immunosorbant assays (ELISAs) and sequencing analyses identified four different scFv antibodies with high sensitivity. Soluble antibody ELISAs identified two highly sensitive scFv antibodies, FvCA3 and FvCA4, with the latter being slightly more sensitive. Three-dimensional modeling revealed that the FvCA4 may hold a better overall structure with CDRH3, CDRL1 and CDRL3 centered in the core region of antibody surface compared with that of other scFvs. Immunofluorescence labeling revealed that the binding of FvCA4 antibody was localized to the cell walls of conidiospores and hyphae of F. verticillioides, confirming the specificity of this antibody for a surface target. This scFv antibody was able to detect the fungal mycelium as low as 10(−2) μg/mL and contaminating mycelium at a quantity of 10(−2) mg/g maize. This is the first report that scFv antibodies derived from phage display have a wide application for rapid and accurate detection and monitoring of fumonisin-producing pathogens in agricultural samples. Molecular Diversity Preservation International (MDPI) 2012-06-07 /pmc/articles/PMC3397510/ /pubmed/22837678 http://dx.doi.org/10.3390/ijms13067038 Text en © 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Hu, Zu-Quan
Liu, Jin-Long
Li, He-Ping
Xing, Shu
Xue, Sheng
Zhang, Jing-Bo
Wang, Jian-Hua
Nölke, Greta
Liao, Yu-Cai
Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display
title Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display
title_full Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display
title_fullStr Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display
title_full_unstemmed Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display
title_short Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display
title_sort generation of a highly reactive chicken-derived single-chain variable fragment against fusarium verticillioides by phage display
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3397510/
https://www.ncbi.nlm.nih.gov/pubmed/22837678
http://dx.doi.org/10.3390/ijms13067038
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