College of Life Science, Shandong Key Laboratory of Agricultural Microbiology, Shandong Agricultural University, Tai'an 271018, China.
College of Life Science, Shandong Key Laboratory of Agricultural Microbiology, Shandong Agricultural University, Tai'an 271018, China.
Food Chem. 2022 Nov 1;393:133317. doi: 10.1016/j.foodchem.2022.133317. Epub 2022 May 26.
Noncompetitive immunoassays for small molecules are generally considered to be more sensitive than competitive ones. In this study, a phage-peptide against immune complex of aflatoxin B1 (AFB1) and nanobody Nb28 was obtained by phage-display technology. The phage-peptide was labeled with peroxidase and used to develop a direct noncompetitive magnetic-chemiluminescent enzyme-linked immunoassay (Nc-MCLEIA) for AFB1. The 50% signal saturation concentration (SC) and limit of detection (LOD) of Nc-MCLEIA for AFB1 were 0.089 and 0.006 ng/mL, respectively. Compared with competitive immunoassays developed by the Nb28, the sensitivity and efficiency of Nc-MCLEIA were greatly improved. The recoveries of AFB1 from spiked corn, rice, flour, peanut, peanut oil and corn oil samples ranged from 83.8% to 119.2% with coefficient of variable under 8.9%. Furthermore, parallel analysis of natural corn, rice and flour samples by Nc-MCLEIA and HPLC (high performance liquid chromatography) proved that the Nc-MCLEIA was reliable.
非竞争免疫分析通常被认为比竞争免疫分析更灵敏。本研究利用噬菌体展示技术获得了一种针对黄曲霉毒素 B1（AFB1）免疫复合物的噬菌体肽和纳米抗体 Nb28。该噬菌体肽被标记过氧化物酶，并用于开发用于 AFB1 的直接非竞争磁化学发光酶联免疫分析（Nc-MCLEIA）。Nc-MCLEIA 检测 AFB1 的 50%信号饱和浓度（SC）和检测限（LOD）分别为 0.089 和 0.006ng/mL。与 Nb28 开发的竞争免疫分析相比，Nc-MCLEIA 的灵敏度和效率得到了极大提高。从玉米、大米、面粉、花生、花生油和玉米油样品中添加 AFB1 的回收率在 83.8%至 119.2%之间，变异系数小于 8.9%。此外，通过 Nc-MCLEIA 和高效液相色谱（HPLC）平行分析天然玉米、大米和面粉样品证明了 Nc-MCLEIA 的可靠性。
