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A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies

Simultaneous aflatoxin (AFB1) and zearalenone (ZEN) contamination in agro-products have become widespread globally and have a toxic superposition effect. In the present study, we describe a highly sensitive and specific dual lateral flow immunochromatographic assay (dual test strip) for rapid and si...

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Autores principales: Wang, Yanan, Wang, Xiaofei, Wang, Shuyun, Fotina, Hanna, Wang, Ziliang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9505352/
https://www.ncbi.nlm.nih.gov/pubmed/36136553
http://dx.doi.org/10.3390/toxins14090615
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author Wang, Yanan
Wang, Xiaofei
Wang, Shuyun
Fotina, Hanna
Wang, Ziliang
author_facet Wang, Yanan
Wang, Xiaofei
Wang, Shuyun
Fotina, Hanna
Wang, Ziliang
author_sort Wang, Yanan
collection PubMed
description Simultaneous aflatoxin (AFB1) and zearalenone (ZEN) contamination in agro-products have become widespread globally and have a toxic superposition effect. In the present study, we describe a highly sensitive and specific dual lateral flow immunochromatographic assay (dual test strip) for rapid and simultaneous detection of AFB1 and ZEN in food and feed samples based on respective monoclonal antibodies (mAbs). Two immunogens AFB1-BSA (an AFB1 and bovine serum albumin (BSA) conjugate) and ZEN-BSA (a ZEN and BSA conjugate) were synthesized in oximation active ester (OAE) and amino glutaraldehyde (AGA). The molecular binding ratio of AFB1:BSA was 8.64:1, and that of ZEN:BSA was 17.2:1, identified by high-resolution mass spectrometry (HRMS) and an ultraviolet spectrometer (UV). The hybridoma cell lines 2A11, 2F6, and 3G2 for AFB1 and 2B6, 4D9 for ZEN were filtered by an indirect non-competitive enzyme-linked immunosorbent assay (inELISA) and an indirect competitive enzyme-linked immunosorbent assay (icELISA), respectively. As AFB1 mAb 2A11 and ZEN mAb 2B6 had the lowest 50% inhibitive concentration (IC50) and cross-reactivity (CR), they were selected for subsequent experiments. By systematically optimizing the preparation condition of gold nanoparticles (AuNPs), AuNPs-labeled mAbs, and detection condition, the visual limit of detection (LOD) of the dual test strip was 1.0 μg/L for AFB1 and 5.0 μg/L for ZEN, whereas that of the test strip reader was 0.23 μg/L for AFB1 and 1.53 μg/L for ZEN. The high reproducibility and stability of the dual test were verified using mycotoxin-spiked samples. The dual test strips were highly specific and sensitive for AFB1 and ZEN, which were validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Thus, the proposed AFB1 and ZEN dual test strip is suitable for rapid and simultaneous detection of AFB1 and ZEN contamination in food and feed samples.
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spelling pubmed-95053522022-09-24 A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies Wang, Yanan Wang, Xiaofei Wang, Shuyun Fotina, Hanna Wang, Ziliang Toxins (Basel) Article Simultaneous aflatoxin (AFB1) and zearalenone (ZEN) contamination in agro-products have become widespread globally and have a toxic superposition effect. In the present study, we describe a highly sensitive and specific dual lateral flow immunochromatographic assay (dual test strip) for rapid and simultaneous detection of AFB1 and ZEN in food and feed samples based on respective monoclonal antibodies (mAbs). Two immunogens AFB1-BSA (an AFB1 and bovine serum albumin (BSA) conjugate) and ZEN-BSA (a ZEN and BSA conjugate) were synthesized in oximation active ester (OAE) and amino glutaraldehyde (AGA). The molecular binding ratio of AFB1:BSA was 8.64:1, and that of ZEN:BSA was 17.2:1, identified by high-resolution mass spectrometry (HRMS) and an ultraviolet spectrometer (UV). The hybridoma cell lines 2A11, 2F6, and 3G2 for AFB1 and 2B6, 4D9 for ZEN were filtered by an indirect non-competitive enzyme-linked immunosorbent assay (inELISA) and an indirect competitive enzyme-linked immunosorbent assay (icELISA), respectively. As AFB1 mAb 2A11 and ZEN mAb 2B6 had the lowest 50% inhibitive concentration (IC50) and cross-reactivity (CR), they were selected for subsequent experiments. By systematically optimizing the preparation condition of gold nanoparticles (AuNPs), AuNPs-labeled mAbs, and detection condition, the visual limit of detection (LOD) of the dual test strip was 1.0 μg/L for AFB1 and 5.0 μg/L for ZEN, whereas that of the test strip reader was 0.23 μg/L for AFB1 and 1.53 μg/L for ZEN. The high reproducibility and stability of the dual test were verified using mycotoxin-spiked samples. The dual test strips were highly specific and sensitive for AFB1 and ZEN, which were validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Thus, the proposed AFB1 and ZEN dual test strip is suitable for rapid and simultaneous detection of AFB1 and ZEN contamination in food and feed samples. MDPI 2022-09-02 /pmc/articles/PMC9505352/ /pubmed/36136553 http://dx.doi.org/10.3390/toxins14090615 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Wang, Yanan
Wang, Xiaofei
Wang, Shuyun
Fotina, Hanna
Wang, Ziliang
A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies
title A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies
title_full A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies
title_fullStr A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies
title_full_unstemmed A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies
title_short A Novel Lateral Flow Immunochromatographic Assay for Rapid and Simultaneous Detection of Aflatoxin B1 and Zearalenone in Food and Feed Samples Based on Highly Sensitive and Specific Monoclonal Antibodies
title_sort novel lateral flow immunochromatographic assay for rapid and simultaneous detection of aflatoxin b1 and zearalenone in food and feed samples based on highly sensitive and specific monoclonal antibodies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9505352/
https://www.ncbi.nlm.nih.gov/pubmed/36136553
http://dx.doi.org/10.3390/toxins14090615
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