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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

1
College of Food Science, Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China
2
Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
3
Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China
*
Authors to whom correspondence should be addressed.
Biomolecules 2019, 9(12), 770; https://doi.org/10.3390/biom9120770
Received: 21 October 2019 / Revised: 17 November 2019 / Accepted: 19 November 2019 / Published: 23 November 2019
(This article belongs to the Section Biochemistry)
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 (IC50) 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. View Full-Text
Keywords: hapten; monoclonal antibody; immunoassay; tylosin; tilmicosin hapten; monoclonal antibody; immunoassay; tylosin; tilmicosin
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MDPI and ACS Style

Huang, J.-X.; Yao, C.-Y.; Yang, J.-Y.; Li, Z.-F.; He, F.; Tian, Y.-X.; Wang, H.; Xu, Z.-L.; Shen, Y.-D. Design of Novel Haptens and Development of Monoclonal Antibody-Based Immunoassays for the Simultaneous Detection of Tylosin and Tilmicosin in Milk and Water Samples. Biomolecules 2019, 9, 770.

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