Sotnikov Dmitriy V, Barshevskaya Lyubov V, Bartosh Anastasia V, Zherdev Anatoly V, Dzantiev Boris B
A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky prospect 33, Moscow 119071, Russia.
Biosensors (Basel). 2022 May 17;12(5):343. doi: 10.3390/bios12050343.
A new scheme of reagents interaction for lateral flow immunoassay (LFIA) is proposed, which combines the features of competitive and sandwich assay and provides highly sensitive detection of low-molecular-weight analytes. Namely, the antigen in the sample interferes with the formation of the antibody (on the membrane)-hapten-protein-antibody (on the nanoparticle-marker) complex, competing with hapten-protein conjugate in both reactions. The proposed scheme was modelled using COPASI software, with a prediction of limit of detection (LOD) decrease by one order of magnitude compared to the standard competitive LFIA. This feature was experimentally confirmed for the detection of chloramphenicol (CAP) in honey. When tested in spiked honey, the visual LOD was 50 ng/mL for the common scheme and 5 ng/mL for the proposed scheme. Instrumental LOD was 300 pg/mL (1.2 µg/kg in conversion per sample weight of honey) in the standard scheme and 20 pg/mL (80 ng/kg in conversion per sample weight of honey) in the proposed scheme.
提出了一种用于侧向流动免疫分析（LFIA）的试剂相互作用新方案，该方案结合了竞争法和夹心分析法的特点，可对低分子量分析物进行高灵敏度检测。具体而言，样品中的抗原会干扰抗体（在膜上）-半抗原-蛋白质-抗体（在纳米颗粒标记物上）复合物的形成，在两个反应中与半抗原-蛋白质偶联物竞争。使用COPASI软件对所提出的方案进行建模，预测检测限（LOD）比标准竞争型LFIA降低一个数量级。这一特性在蜂蜜中氯霉素（CAP）检测中得到了实验证实。在加标蜂蜜中进行测试时，普通方案的视觉LOD为50 ng/mL，所提出方案的视觉LOD为5 ng/mL。标准方案的仪器LOD为300 pg/mL（按蜂蜜样品重量换算为1.2 µg/kg），所提出方案的仪器LOD为20 pg/mL（按蜂蜜样品重量换算为80 ng/kg）。
