title: Monoclonal antibody production and the development of an indirect competitive enzyme-linked immunosorbent assay for screening spiramycin in milk.
abstract: 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 μg/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 μg/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.
authors.affiliation: College of Veterinary Medicine, China Agricultural University , Beijing 100193, People's Republic of China.
authors.forename: Wenxiao
authors.initials: W
authors.lastname: Jiang
authors.forename: Huiyan
authors.initials: H
authors.lastname: Zhang
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Li
authors.forename: Xinxin
authors.initials: X
authors.lastname: Liu
authors.forename: Suxia
authors.initials: S
authors.lastname: Zhang
authors.forename: Weimin
authors.initials: W
authors.lastname: Shi
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis; D002417:Cattle; D004350:Drug Residues / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D005260:Female; D005506:Food Contamination / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry*; D015572:Spiramycin / Q000032:analysis*
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D015572:Spiramycin