A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia.
Biosensors (Basel). 2022 Dec 14;12(12):1166. doi: 10.3390/bios12121166.
The work considers a combination of three enhancing approaches for immunochromatographic assay (ICA) and the integration of their impacts into changes of the limit of detection (LOD). Human fatty acid binding protein (FABP), an early biomarker of acute myocardial infarction, was the target analyte. Starting from the common ICA protocol with an LOD equal to 11.2 ng/mL, three approaches were realized: (1) replacement of spherical gold nanoparticles with gold nanoflowers having a branched surface (20-fold lowering the LOD); (2) enhanced labeling of immune complexes via nanoparticle aggregates (15-fold lowering); (3) in-situ growth of bound nanoparticles by reduction of gold salts (3-fold lowering). Single and combined implementations of these approaches have been studied. It has been shown that the LOD decrease for combined approaches is close to the multiplied contribution of each of them. The final LOD for FABP was 0.05 ng/mL, which is 220 times lower than the LOD for the common ICA protocol. The efficiency of the enhanced ICA with three combined approaches was confirmed by testing human serum samples for FABP presence and content. The development presents a new efficient technique for rapid sensitive detection of FABP for medical diagnostics. Moreover, the demonstrated multiple enhancements could be applied for various demanded analytes.
该工作考虑了三种增强免疫层析分析（ICA）的方法，并将它们的影响整合到检测限（LOD）的变化中。人脂肪酸结合蛋白（FABP）是急性心肌梗死的早期生物标志物，是目标分析物。从 LOD 等于 11.2ng/mL 的常见 ICA 方案开始，实现了三种方法：（1）用具有分支表面的金纳米花替代球形金纳米粒子（将 LOD 降低 20 倍）；（2）通过纳米粒子聚集体增强免疫复合物的标记（降低 15 倍）；（3）通过还原金盐原位生长结合的纳米粒子（降低 3 倍）。研究了这些方法的单独和组合实施情况。结果表明，组合方法的 LOD 降低接近于它们各自贡献的乘积。FABP 的最终 LOD 为 0.05ng/mL，比常见的 ICA 方案的 LOD 低 220 倍。通过测试人血清样本中 FABP 的存在和含量，验证了具有三种组合方法的增强 ICA 的效率。该研究提出了一种用于医学诊断的快速灵敏检测 FABP 的新技术。此外，所展示的多种增强方法可应用于各种所需的分析物。
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