A Class-Selective Immunoassay for Sulfonamides Residue Detection in Milk Using a Superior Polyclonal Antibody with Broad Specificity and Highly Uniform Affinity

Molecules

Kai Wen¹  Zhanhui Wang¹  Weimin Shi¹  Yanxin Cao¹  Chao Li¹  Xiangshu Luo¹  Chenglong Li¹  Huijuan Yang¹  Xuezhi Yu¹  Suxia Zhang¹  Weiyu Wang¹  Xiao Liang²  Yonghan Li³ 

[1] Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory of Food Quality and Safety, Beijing 100193, China;College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China;Henan Animal Health Supervision Institute, Zhengzhou 450008, China;

关键词: milk safety;    multi-sulfonamides;    novel hapten;    polyclonal antibody;    broad specificity;    highly uniform affinity;   

DOI  :  10.3390/molecules24030443

来源: DOAJ

【 摘 要 】

The development of multianalyte immunoassays with an emphasis on food safety has attracted increasing interest, due to its high target throughput, short detection time, reduced sample consumption, and low overall cost. In this study, a superior polyclonal antibody (pAb) against sulfonamides (SAs) was raised by using a bioconjugate of bovine serum albumin with a rationally designed hapten 4-[(4-aminophenyl) sulfonyl-amino]-2-methoxybenzoic acid (SA10-X). The results showed that the pAb could recognize 19 SAs with 50% inhibition (IC₅₀) below 100 µg L⁻¹ and a recognition profile for SAs containing, either a five-atom ring or a six-atom ring, with highly uniform affinity. A three-dimensional quantitative structure-activity relationship analysis indicated that the electrostatic features of SAs play a considerably important role, during recognition with pAb than stereochemical effects. Skimmed milk samples were directly diluted five times before analysis. After optimization, the limit of detection for sulfamonomethoxine, sulfamethoxazole, sulfaquinoxaline, sulfadimethoxine, and sulfamethazine were 1.00, 1.25, 2.95, 3.35, and 6.10 µg L⁻¹, respectively. The average recoveries for these 5 SAs were 72.0⁻107.5% with coefficients of variation less than 14.1%. The established method, based on pAb, with broad specificity and uniform affinity, offered a simple, sensitive, and high-throughput screening tool for the detection of multi-SAs in milk samples.

【 授权许可】

Unknown