建立一种基于单克隆抗体的免疫层析法用于检测卡马西平和卡马西平 10,11-环氧化物。

Development of a monoclonal antibody-based immunochromatographic assay for the detection of carbamazepine and carbamazepine-10, 11-epoxide.

机构信息

State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, People's Republic of China; International Joint Research Laboratory for Biointerface and Biodetection, School of Food Science and Technology, Jiangnan University, Wuxi, People's Republic of China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Mar 15;1141:122036. doi: 10.1016/j.jchromb.2020.122036. Epub 2020 Feb 15.

DOI:10.1016/j.jchromb.2020.122036

PMID:32109745

Abstract

In this study, haptens were designed to produce highly sensitive and specific monoclonal antibodies (mAb) against carbamazepine (CBZ) and its metabolite carbamazepine-10, 11-epoxide (CBZ-EP). According to the results of our competitive enzyme-linked immunosorbent assay (ic-ELISA), the half-maximum inhibitory concentration values for anti-CBZ and anti-CBZ-EP mAb were 0.18 and 0.59 ng/mL, respectively. An immunochromatographic assay (ICA) was developed for the determination of CBZ and CBZ-EP concentrations. This method can provide visible limits of detection ranging from 0.25 to 1 ng/mL, and cut-off limits ranging from 5 to 10 ng/mL, and takes 10 min to evaluate with the naked eye. Importantly, these observations were consistent with those obtained by ic-ELISA and liquid chromatography-mass spectrometry. The ICA assay represented a reliable, fast, and high-throughput method for the determination of CBZ and CBZ-EP in serum samples.

摘要

在这项研究中，设计了半抗原以产生针对卡马西平（CBZ）及其代谢物卡马西平-10,11-环氧化物（CBZ-EP）的高度敏感和特异性单克隆抗体（mAb）。根据我们的竞争性酶联免疫吸附测定（ic-ELISA）的结果，抗 CBZ 和抗 CBZ-EP mAb 的半最大抑制浓度值分别为 0.18 和 0.59 ng/mL。开发了免疫层析测定法（ICA）来测定 CBZ 和 CBZ-EP 的浓度。该方法可以提供从 0.25 到 1 ng/mL 的可见检测限，以及从 5 到 10 ng/mL 的截止限，并且可以在 10 分钟内用肉眼评估。重要的是，这些观察结果与 ic-ELISA 和液相色谱-质谱法的结果一致。ICA 测定法是一种可靠，快速且高通量的方法，可用于测定血清样品中的 CBZ 和 CBZ-EP。