Department of Medical Diagnostics, Centre for Advanced Materials and Technologies CEZAMAT, Warsaw University of Technology, Poleczki 19, 02-822, Warsaw, Poland; Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Stanisława Noakowskiego 3, 00-664, Warsaw, Poland; Section of Inorganic and Analytical Chemistry, Faculty of Chemistry, University of Warsaw, Ludwika Pasteura 1, 02-093, Warsaw, Poland.
Department of Medical Diagnostics, Centre for Advanced Materials and Technologies CEZAMAT, Warsaw University of Technology, Poleczki 19, 02-822, Warsaw, Poland; Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Stanisława Noakowskiego 3, 00-664, Warsaw, Poland.
Biosens Bioelectron. 2023 Oct 1;237:115511. doi: 10.1016/j.bios.2023.115511. Epub 2023 Jul 6.
Scientific interest in the investigation and application of multifunctional nanomaterials in medical diagnostics has been increasing. The employment of magnetocatalytic immunoconjugates as both analyte-capturing agents and enzyme-like catalytic labels may enable rapid preconcentration and determination of relevant antigens. In this work, we synthesized and comprehensively characterized two types of noble metal-decorated magnetic nanocubes (MDMCs) which were subsequently applied in the one-step, sandwich nanozyme-linked immunosorbent assay (NLISA). Magnetic cores allow for rapid separation from complex samples of biological origin. The catalytically active shell composed of Au-decorated Pt or Ru can effectively mimic the activity of horseradish peroxididase, retaining at the same time the ability to form stable bioconstructs through self-assembly of thiolated ligands. As a result, hybrid multifunctional nanoparticles were synthesized and used to detect C-reactive protein (CRP) in serum samples. We have also paid considerable attention to the mechanistic studies of the formation of sandwich immunocomplexes with nanoparticle labels by means of immunoenzymatic methods and surface plasmon resonance. Analytical parameters of the Pt-MDMCs-labeled NLISA (detection limit LOD = 0.336 ng mL, recovery = 98.0%, linear response window covering two logarithmic units) turned out to be superior to the classical, one-step ELISA based on a horseradish peroxidase. In addition, our method offers further possibility of sensitivity adjustment by changing the parameters of magnetic preconcentration, together with good long-term stability of MDMCs conjugates and their good resistance to common interferences. We believe that the proposed simple synthetic protocol will guide a new approach to applying metal-decorated magnetic nanozymes as versatile and multifunctional labels for application in subsequent pre-analytical analyte concentration and immunoassays towards clinical applications.
科学研究越来越关注多功能纳米材料在医学诊断中的应用。将磁催化免疫缀合物用作分析物捕获剂和类酶催化标签，可以实现相关抗原的快速预浓缩和测定。在这项工作中，我们合成并全面表征了两种类型的贵金属修饰磁性纳米立方（MDMC），随后将其应用于一步夹心纳米酶联免疫吸附测定（NLISA）中。磁性核可实现与生物来源的复杂样品的快速分离。由 Au 修饰的 Pt 或 Ru 组成的催化活性壳可以有效地模拟辣根过氧化物酶的活性，同时保持通过硫醇配体的自组装形成稳定生物构建体的能力。结果，合成了杂化多功能纳米粒子，并用于检测血清样品中的 C 反应蛋白（CRP）。我们还非常关注通过免疫酶方法和表面等离子体共振研究纳米粒子标签形成夹心免疫复合物的形成机制。Pt-MDMC 标记 NLISA 的分析参数（检测限 LOD = 0.336 ng mL，回收率 = 98.0%，线性响应窗口覆盖两个对数单位）优于基于辣根过氧化物酶的经典一步 ELISA。此外，通过改变磁预浓缩的参数，我们的方法还可以进一步调整灵敏度，并且 MDMC 缀合物具有良好的长期稳定性和对常见干扰的良好抵抗力。我们相信，所提出的简单合成方案将为应用金属修饰的磁性纳米酶作为多功能和多功能标签提供新的方法，用于随后的预分析物浓缩和免疫分析，以实现临床应用。
