Faculty of Pharmaceutical Sciences, Prince of Songkla University, Songkhla, Thailand.
School of Pharmacy, Walailak University, Nakhon Si Thammarat, Thailand.
Phytochem Anal. 2023 Jun;34(4):421-430. doi: 10.1002/pca.3223. Epub 2023 Mar 23.
Miroestrol (Mi) and deoxymiroestrol (Dmi) are trace, yet potent, phytooestrogens found in white Kwao Krua [Pueraria candollei var. mirifica (Airy Shaw & Suvat.) Niyomdham, PM]. However, the analysis of these substances is difficult because of complex matrix effects and their various analogues. In addition, alteration in the cross-reactivity of a gold nanoparticle (AuNP)-based immunochromatographic assay (ICA) resulting from the electrostatic adsorption between antibodies and AuNPs has not yet been evaluated.
This study aims to develop, characterise, and validate ICA with a monoclonal antibody exhibiting similar reactivity against Mi and Dmi (MD-mAb).
The ICA performance was validated for cross-reactivity and performance in comparison with those of indirect competitive enzyme-linked immunosorbent assays (icELISAs) with MD-mAb and mAb exhibiting specificity against Mi (Mi-mAb).
The ICA showed a limit of detection (LOD) at 1 and 16 μg/mL for Mi and Dmi, respectively. The cross-reactivity of the ICA with Dmi was lower (6.25%) than that observed with the icELISA (120%). Cross-reactivity of ICA against other compounds of the PM was also correlated with those of icELISA; no false-positive/negative results were observed. The repeatability and reproducibility of the ICA were confirmed. The results obtained using ICA in samples of PM are correlated with the concentrations determined through icELISAs.
An ICA with MD-mAb was constructed and validated. However, direct conjugation via the electrostatic adsorption of mAb-AuNPs was expected to alter the cross-reactivity of ICA, especially that of the analyte analogue Dmi.
米罗昔芬（Mi）和去氧米罗昔芬（Dmi）是微量但强效的植物雌激素，存在于白 Kwao Krua [三叶野葛（ Pueraria candollei var. mirifica （Airy Shaw & Suvat.）Niyomdham，PM）]中。然而，由于基质效应复杂以及存在各种类似物，这些物质的分析较为困难。此外，金纳米颗粒（AuNP）基于免疫层析测定（ICA）的交叉反应性因抗体和 AuNP 之间的静电吸附而发生变化，尚未得到评估。
本研究旨在开发、表征和验证基于单克隆抗体的 ICA，该抗体对 Mi 和 Dmi（MD-mAb）具有相似的反应性。
对 ICA 的交叉反应性和性能进行了验证，并与 MD-mAb 和针对 Mi（Mi-mAb）具有特异性的 mAb 进行间接竞争酶联免疫吸附测定（icELISA）进行了比较。
ICA 的 Mi 和 Dmi 的检测限（LOD）分别为 1 和 16μg/mL。ICA 对 Dmi 的交叉反应性较低（6.25%），而 icELISA 则为 120%。ICA 对 PM 中其他化合物的交叉反应性也与 icELISA 相关；未观察到假阳性/阴性结果。ICA 的重复性和再现性得到了验证。在 PM 样本中使用 ICA 获得的结果与通过 icELISAs 确定的浓度相关。
构建并验证了基于 MD-mAb 的 ICA。然而，通过 mAb-AuNP 的静电吸附直接偶联预计会改变 ICA 的交叉反应性，尤其是分析物类似物 Dmi 的交叉反应性。
