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

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 IC50 = 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.