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

Autor:	Fan He, Zhenfeng Li, Yu-Dong Shen, Hong Wang, Zhen-Lin Xu, Jian-Xin Huang, Chan-Yuan Yao, Yuanxin Tian, Jin-Yi Yang
Rok vydání:	2019
Předmět:	medicine.drug_class animal diseases Enzyme-Linked Immunosorbent Assay Food Contamination Fresh Water 010501 environmental sciences Tylosin tilmicosin Monoclonal antibody 01 natural sciences Biochemistry High-performance liquid chromatography Article chemistry.chemical_compound fluids and secretions medicine Animals immunoassay Tilmicosin Molecular Biology Drug toxicity 0105 earth and related environmental sciences Food hygiene tylosin Chromatography medicine.diagnostic_test 010401 analytical chemistry Antibodies Monoclonal Anti-Bacterial Agents 0104 chemical sciences Milk chemistry monoclonal antibody hapten Immunoassay Cattle Haptens Hapten
Zdroj:	Biomolecules Volume 9 Issue 12
ISSN:	2218-273X
DOI:	10.3390/biom9120770
Popis:	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.
Databáze:	OpenAIRE
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