A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia.
A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia.
Talanta. 2017 Dec 1;175:77-81. doi: 10.1016/j.talanta.2017.07.027. Epub 2017 Jul 11.
The scheme of immunochromatographic competitive analysis with "external" specific antibodies is offered for the first time. The test strip includes all the necessary components for analysis (membranes, protein antigen conjugates, colored nanoparticles), except specific free unconjugated antibodies that are transferred to the buffer used for sample dilution. Thus, the concentrations of the antibodies and markers can be chosen independently, and dilution of the sample with buffer (a routine procedure for any immunochromatographic test) ensures pre-incubation of the antibodies with the sample. Both factors increase the sensitivity of the test. The proposed scheme has been proven to be effective for the determination of deoxynivalenol and T-2 toxin; detection limits of 500pg/mL and 50pg/mL, respectively, were achieved. This is an order of magnitude lower than the limits of detection of traditional tests using the same reagents. The analysis was conducted in water-organic extracts (20% methanol); the duration of the analysis is 10min (it's the same time as for a traditional test). The proposed approach is universal, and it can be used to detect a variety of compounds.
本文首次提出了基于“外部”特异性抗体的免疫层析竞争分析方案。该测试条包含分析所需的所有组件(膜、蛋白抗原缀合物、有色纳米颗粒),但不包括特异性游离未缀合抗体,后者被转移到用于稀释样品的缓冲液中。因此,可以独立选择抗体和标记物的浓度,并且用缓冲液稀释样品(这是任何免疫层析测试的常规程序)可以确保抗体与样品的预孵育。这两个因素都提高了测试的灵敏度。该方案已被证明可有效用于测定脱氧雪腐镰刀菌烯醇和 T-2 毒素,检测限分别为 500pg/mL 和 50pg/mL,比使用相同试剂的传统检测方法的检测限低一个数量级。该分析在水-有机溶剂提取物(20%甲醇)中进行;分析时间为 10 分钟(与传统测试相同)。所提出的方法是通用的,可以用于检测多种化合物。