A monoclonal antibody-based enzyme-linked immunosorbent assay for the determination of chlorogenic acid in honeysuckle
The edible flower buds of honeysuckle are traditionally used as herbal medicine or food additives in China. Chlorogenic acid (CGA) serves as the quality control marker of honeysuckle for its high content and antipyretic property. In this paper, a specific monoclonal antibody (mAb2E2) against CGA was produced. After optimization, mAb2E2-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed. The concentrations of CGA producing 50% inhibition and the calibration range of the icELISA were 0.39 and 0.10–1.51 ng/mL, respectively. The icELISA had cross-reactivity with 3,5-Dicaffeoylquinic acid (17.53%), and the cross-reactivity levels with other analogs were all below 5%. The average recoveries obtained by standard CGA addition to honeysuckle samples were from 88.4 to 104.8%. The icELISA was applied to CGA detection in different honeysuckle samples and the results were confirmed by high-performance liquid chromatography analysis. The correlation coefficient between the two assays was 0.97. The proposed icELISA provides a feasible analytical method for highly sensitive and specific, simple, fast, and high-throughput determination of CGA in honeysuckle samples.