Crystarose 4B偶联黄曲霉毒素单克隆抗体方法优化及其应用

Optimization of Crystarose 4B Coupling Aflatoxin Monoclonal Antibody and Its Application

对Crystarose 4B偶联黄曲霉毒素单克隆抗体的方法进行优化,制备了黄曲霉毒素B1,B2,G1,G2免疫亲和柱。采用甲醇-水为提取溶剂,用均质机将样品的粉碎和毒素提取一步完成,免疫亲和柱对提取液进行净化、富集,收集洗脱液,在UPLC-MS/MS多反应监测(MRM)模式下进行了定性和定量分析。实验结果表明,该偶联方法稳定,亲和柱回收率在94. 4%～107. 1%之间,相对标准偏差为1. 5%～4. 9%,重复性、再现性和稳定性良好,柱容量可满足国标检测要求。采用所建立的方法检测花生、玉米和小麦实际样品中的黄曲霉毒素,检出率分别为15%、10%和10%。

The method of coupling Crystarose 4B to Aflatoxin monoclonal antibody was optimized,and Aflatoxin B1,B2,G1,G2 immunoaffinity column was prepared.Sample preparation was simplified and utilized with a methanol-water solution in a one-step grinding extraction step.The extract was then purified using the home-made immunoaffinity column followed by UPLC-MS/MS for quantification.The results showed that the coupling method was stable,the recoveries ranged from 94.4% to 107.1%,with the relative standard deviations (RSDs) of 1.5%~ 4.9%.The immunoaffinity column demonstrated considerable repeatability,reproducibility,and stability,and the column capacity can meet the requirements of the national standard.Then the method was applied in the detection of peanut,corn and wheat,with detection rates of 15%,10% and 10%,respectively.