Bartosh Anastasiya V, Sotnikov Dmitriy V, Zherdev Anatoly V, Dzantiev Boris B
A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia.
Micromachines (Basel). 2023 Jan 28;14(2):333. doi: 10.3390/mi14020333.
Changes in the limits of detection (LODs) for a multiplex lateral flow immunoassay (LFIA) caused by different locations of the binding zone on the test strips were studied. Due to the non-equilibrium conditions of the immune reactions in LFIAs, their analytical parameters are susceptible to the binding constants of antigen-antibody reactions and assay duration. Consequently, the integration of several tests into one multiplex assay can cause a significant worsening of the sensitivity. In this study, we propose a simple methodology for the determination of the best arrangement of binding zones, which takes into account the binding constants for immunoreagents. LFIAs of four mycotoxins, namely, aflatoxin B1, deoxynivalenol, T-2 toxin, and ochratoxin A, were integrated into a multiplex test strip. An enzyme-linked immunosorbent assay was applied to determine the equilibrium and kinetic constants of the immunoreactants for each analyte. It was found that the arrangement of binding zones with a descending order of the equilibrium association constants was optimal and provided both lower detection limits and a more uniform coloration. The selected position of the binding zones allowed decreasing the LODs down to 2 and 27 times for ochratoxin A and deoxynivalenol, respectively. The proposed approach can be applied to multiplex LFIAs for different analytes.
研究了由试纸条上结合区不同位置引起的多重侧向流动免疫分析（LFIA）检测限（LOD）的变化。由于LFIA中免疫反应的非平衡条件，其分析参数易受抗原-抗体反应的结合常数和检测持续时间的影响。因此，将多个检测整合到一个多重检测中可能会导致灵敏度显著下降。在本研究中，我们提出了一种简单的方法来确定结合区的最佳排列方式，该方法考虑了免疫试剂的结合常数。将四种霉菌毒素，即黄曲霉毒素B1、脱氧雪腐镰刀菌烯醇、T-2毒素和赭曲霉毒素A的LFIA整合到一个多重试纸条中。采用酶联免疫吸附测定法测定每种分析物免疫反应物的平衡常数和动力学常数。结果发现，按平衡缔合常数降序排列结合区是最佳的，并且能提供更低的检测限和更均匀的显色。结合区的选定位置分别使赭曲霉毒素A和脱氧雪腐镰刀菌烯醇的检测限降低至2倍和27倍。所提出的方法可应用于不同分析物的多重LFIA。
