期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 61, 期 46, 页码 10925-10931出版社
AMER CHEMICAL SOC
DOI: 10.1021/jf404027b
关键词
spiramycin; monoclonal antibody; immunoassay; residue; milk
资金
- National Natural Science Foundation of China [31372475]
- Trans-Century Training Program Foundation for the Talents by the Ministry of Education [NCET-12-0529]
- National Science & Technology Pillar Program during the twelfth Five-Year Plan Period [2011BAK10B01, 2012BAK17B00]
To monitor spiramycin (SP) residue in milk, a monoclonal antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed. This study described the preparation of three immunogens and the production of a high-affinity mAb. After optimization, the 50% inhibition concentration (IC50) for the developed icELISA was estimated as 0.97 ng/mL in the assay buffer, and the limit of detection and limit of quantitation were 2.51 and 4.40 mu g/L in the milk matrix. The newly developed assay demonstrated negligible cross-reactivity with 15 other macrolide antibiotics, but not with kitasamycin (23.4%). The mean recoveries ranged from 81 to 103% for the spiked samples (5, 10, and 50 mu g/L), and the coefficient of variation ranged from 5.4 to 9.6%. The icELISA was validated by LC-MS/MS method, and all results demonstrated that it was a suitable screening method for detecting SP residue in milk without requiring a cleanup process.
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