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一种用于鉴定地沟油中辣椒素同系物的单克隆抗体及其应用。

A monoclonal antibody for identifying capsaicin congeners in illegal cooking oil and its applications.

作者信息

Wu Yuxiang, Liu Jie, Yu Jinzhi, Zhuang Jinqiu, Ma Fengyun, Tan Jing, Shen Zhiqiang

机构信息

School of Agricultural Engineering and Food Science, Shandong University of Technology, No. 266 Xincun Xilu, Zibo, 255049, Shandong Province, China.

Shandong Lvdu Biotechnology Co., Ltd., Shandong, 256600, China.

出版信息

Talanta. 2022 Dec 1;250:123686. doi: 10.1016/j.talanta.2022.123686. Epub 2022 Jun 13.

Abstract

In this work, we studied the preparation of a high-affinity antibody and its immunochromatographic applications to simultaneously identify capsaicin(LJJ), dihydrocapsaicin(HLJ), nordihydrocapsaicin, homodihydrocapsaicin, and other congeners in illegal cooking oil. We used dihydrocapsaicin hapten-conjugated carrier protein BSA as the immunogen according to the formaldehyde method, and conjugated capsaicin and OVA as the coated detection antigen according to the formaldehyde method. We subsequently screened and cloned a hybridoma cell line 2B3 with the highest affinity, which could stably secrete monoclonal antibodies against compounds in the capsaicin family. We then established a capsaicin indirect ELISA standard curve, which was fitted using the linear regression equation R = 0.9987, curve y = -2.3x + 0.2, and IC = 0.2 ng/mL. The cross-reaction rate for capsaicin was 100%, 116% for dihydrocapsaicin, 88% for homodihydrocapsaicin, and 94% for nordihydrocapsaicin. In the second application, we established a simple and accurate sample pretreatment method and a quantum dot-labeled test strip to quickly and quantitatively detect capsaicin family compounds in illegal cooking oil in 8 min. The average recovery rates for each spiked concentration were between 75% and 107.8%, and the coefficient of variation values for each spiked concentration were less than 15%. The high-affinity antibody we identified could simultaneously identify capsaicin, dihydrocapsaicin, nordihydrocapsaicin, homodihydrocapsaicin, and other congeners in illegal cooking oil, and the antibody could be quickly and accurately applied for the qualitative and quantitative detection of capsaicin family residues in illegal cooking oil.

摘要

在本研究中,我们探究了高亲和力抗体的制备及其免疫色谱应用,以同时鉴定非法食用油中的辣椒素(LJJ)、二氢辣椒素(HLJ)、去甲二氢辣椒素、高二氢辣椒素及其他同系物。我们根据甲醛法,使用二氢辣椒素半抗原偶联载体蛋白牛血清白蛋白作为免疫原,并根据甲醛法将辣椒素与卵清蛋白偶联作为包被检测抗原。随后,我们筛选并克隆了亲和力最高的杂交瘤细胞系2B3,其能够稳定分泌针对辣椒素家族化合物的单克隆抗体。接着,我们建立了辣椒素间接ELISA标准曲线,通过线性回归方程R = 0.9987拟合,曲线为y = -2.3x + 0.2,IC = 0.2 ng/mL。辣椒素的交叉反应率为100%,二氢辣椒素为116%,高二氢辣椒素为88%,去甲二氢辣椒素为94%。在第二项应用中,我们建立了一种简单准确的样品预处理方法和量子点标记试纸条,可在8分钟内快速定量检测非法食用油中的辣椒素家族化合物。各加标浓度的平均回收率在75%至107.8%之间,各加标浓度的变异系数值小于15%。我们鉴定出的高亲和力抗体能够同时鉴定非法食用油中的辣椒素、二氢辣椒素、去甲二氢辣椒素、高二氢辣椒素及其他同系物,该抗体可快速准确地应用于非法食用油中辣椒素家族残留物的定性和定量检测。

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