Journal
BIOMOLECULES
Volume 9, Issue 12, Pages -Publisher
MDPI
DOI: 10.3390/biom9120770
Keywords
hapten; monoclonal antibody; immunoassay; tylosin; tilmicosin
Categories
Funding
- National Key Research and Development of China [2018YFC1602904]
- National Natural Science Foundation of China [31871887]
- Science and Technology Planning Project of Guangzhou [201704020082]
- Key Scientific Research Projects of Guangdong Provincial Universities and Colleges [2018KZDXM011]
- Guangdong Provincial Natural Science Foundation [2018B030314005]
- Science and Technology Planning Project of the Guangxi [2017AB47020]
- Graduate Student Overseas Study Program of South China Agricultural University [2019LHPY003]
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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.
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