Reichmuth Andreas M, Kübrich Katharina, Blickenstorfer Yves, Frutiger Andreas, Momotenko Dmitry, Gatterdam Volker, Treindl Fridolin, Fattinger Christof, Vörös János
Laboratory of Biosensors and Bioelectronics, Institute of Biomedical Engineering, ETH Zurich, 8092 Zurich, Switzerland.
Roche Pharma Research and Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
ACS Sens. 2021 Mar 26;6(3):1067-1076. doi: 10.1021/acssensors.0c02346. Epub 2021 Feb 25.
In vitro diagnostics relies on the quantification of minute amounts of a specific biomolecule, called biomarker, from a biological sample. The majority of clinically relevant biomarkers for conditions beyond infectious diseases are detected by means of binding assays, where target biomarkers bind to a solid phase and are detected by biochemical or physical means. Nonspecifically bound biomolecules, the main source of variation in such assays, need to be washed away in a laborious process, restricting the development of widespread point-of-care diagnostics. Here, we show that a diffractometric assay provides a new, label-free possibility to investigate complex samples, such as blood plasma. A coherently arranged sub-micron pattern, that is, a peptide mologram, is created to demonstrate the insensitivity of this diffractometric assay to the unwanted masking effect of nonspecific interactions. In addition, using an array of low-affinity binders, we also demonstrate the feasibility of molecular profiling of blood plasma in real time and show that individual patients can be differentiated based on the binding kinetics of circulating proteins.
体外诊断依赖于从生物样本中对微量特定生物分子（称为生物标志物）进行定量分析。对于传染病以外的病症，大多数临床相关生物标志物是通过结合测定法检测的，其中目标生物标志物与固相结合，并通过生化或物理手段进行检测。非特异性结合的生物分子是此类测定中变异的主要来源，需要通过繁琐的过程将其洗去，这限制了广泛的即时诊断的发展。在此，我们表明衍射测定法为研究复杂样本（如血浆）提供了一种新的、无标记的可能性。创建了一种相干排列的亚微米图案，即肽全息图，以证明这种衍射测定法对非特异性相互作用的有害掩蔽效应不敏感。此外，使用一系列低亲和力结合剂，我们还证明了实时对血浆进行分子谱分析的可行性，并表明可以根据循环蛋白的结合动力学区分个体患者。
