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Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples

In this work, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was established to detect tylosin and tilmicosin in milk and water samples. A sensitive and specific monoclonal antibody was prepared by rational designed hapten, which was achieved by directly...

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Autores principales: Huang, Jian-Xin, Yao, Chan-Yuan, Yang, Jin-Yi, Li, Zhen-Feng, He, Fan, Tian, Yuan-Xin, Wang, Hong, Xu, Zhen-Lin, Shen, Yu-Dong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6995535/
https://www.ncbi.nlm.nih.gov/pubmed/31771142
http://dx.doi.org/10.3390/biom9120770
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author Huang, Jian-Xin
Yao, Chan-Yuan
Yang, Jin-Yi
Li, Zhen-Feng
He, Fan
Tian, Yuan-Xin
Wang, Hong
Xu, Zhen-Lin
Shen, Yu-Dong
author_facet Huang, Jian-Xin
Yao, Chan-Yuan
Yang, Jin-Yi
Li, Zhen-Feng
He, Fan
Tian, Yuan-Xin
Wang, Hong
Xu, Zhen-Lin
Shen, Yu-Dong
author_sort Huang, Jian-Xin
collection PubMed
description In this work, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was established to detect tylosin and tilmicosin in milk and water samples. A sensitive and specific monoclonal antibody was prepared by rational designed hapten, which was achieved by directly oxidizing the aldehyde group on the side chain of tylosin to the carboxyl group. Under the optimized conditions, the linear range of icELISA for tylosin and tilmicosin were 1.3 to 17.7 ng/mL and 2.0 to 47.4 ng/mL, with half-maximal inhibition concentration (IC(50)) values of 4.7 and 9.6 ng/mL, respectively. The cross-reactivity with other analogues of icELISA was less than 0.1%. The average recoveries of icELISA for tylosin and tilmicosin ranged from 76.4% to 109.5% in milk and water samples. Besides, the detection results of icELISA showed good correlations with HPLC-MS/MS. The proposed icELISA was satisfied for rapid and specific screening of tylosin and tilmicosin residues in milk and water samples.
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spelling pubmed-69955352020-02-13 Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples Huang, Jian-Xin Yao, Chan-Yuan Yang, Jin-Yi Li, Zhen-Feng He, Fan Tian, Yuan-Xin Wang, Hong Xu, Zhen-Lin Shen, Yu-Dong Biomolecules Article In this work, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was established to detect tylosin and tilmicosin in milk and water samples. A sensitive and specific monoclonal antibody was prepared by rational designed hapten, which was achieved by directly oxidizing the aldehyde group on the side chain of tylosin to the carboxyl group. Under the optimized conditions, the linear range of icELISA for tylosin and tilmicosin were 1.3 to 17.7 ng/mL and 2.0 to 47.4 ng/mL, with half-maximal inhibition concentration (IC(50)) values of 4.7 and 9.6 ng/mL, respectively. The cross-reactivity with other analogues of icELISA was less than 0.1%. The average recoveries of icELISA for tylosin and tilmicosin ranged from 76.4% to 109.5% in milk and water samples. Besides, the detection results of icELISA showed good correlations with HPLC-MS/MS. The proposed icELISA was satisfied for rapid and specific screening of tylosin and tilmicosin residues in milk and water samples. MDPI 2019-11-23 /pmc/articles/PMC6995535/ /pubmed/31771142 http://dx.doi.org/10.3390/biom9120770 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Huang, Jian-Xin
Yao, Chan-Yuan
Yang, Jin-Yi
Li, Zhen-Feng
He, Fan
Tian, Yuan-Xin
Wang, Hong
Xu, Zhen-Lin
Shen, Yu-Dong
Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples
title Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples
title_full Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples
title_fullStr Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples
title_full_unstemmed Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples
title_short Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples
title_sort design of novel haptens and development of monoclonal antibody-based immunoassays for the simultaneous detection of tylosin and tilmicosin in milk and water samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6995535/
https://www.ncbi.nlm.nih.gov/pubmed/31771142
http://dx.doi.org/10.3390/biom9120770
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