Zeidan Effat, Li Siqi, Zhou Zhiguo, Miller Jennifer, Sandros Marinella G
Department of Nanoscience, University of North Carolina at Greensboro, Greensboro, NC 27455, USA.
Luna Innovations Incorporated, Danville, VA 24541, USA.
Sci Rep. 2016 Oct 31;6:36348. doi: 10.1038/srep36348.
The clinical assessment of multiple organ dysfunctions at early stages is recognized to be an important factor in prompting definitive treatment decisions that prevent irreversible organ damage. In this article, we propose a real-time, label-free, and multiplex nanoenhanced SPRi platform to quantitatively assess two biomarkers, kidney injury molecule (KIM-1) and high mobility group box-1 (HMGB-1) simultaneously in buffer. Our work involves three major contributions in the design of the immunosensor: (1) we applied site-specific immobilization of antibodies to the solid surface that avoids loss of biological activity caused by covalent attachment; (2) we constructed a well-blocked sensor surface that exhibits minimal non-specific adsorption for singleplex measurements of each biomarker in buffer; and (3) we adopted a sandwich assay that implements functionalized quantum dots (NanoEnhancers) as signal amplifiers to achieve a sensitivity level of 5 pg/mL for KIM-1 and HMGB-1 in buffer. We foresee great potential and success in extending this multiplex and ultra-sensitive platform to assess a variety of other emerging clinical biomarkers at low concentrations and in complex matrices.
早期对多器官功能障碍进行临床评估被认为是促使做出预防不可逆器官损伤的确定性治疗决策的重要因素。在本文中，我们提出了一种实时、无标记且可多重检测的纳米增强表面等离子体共振成像（SPRi）平台，用于在缓冲液中同时定量评估两种生物标志物，即肾损伤分子（KIM-1）和高迁移率族蛋白B1（HMGB-1）。我们的工作在免疫传感器设计方面有三大贡献：（1）我们将抗体位点特异性固定在固体表面，避免了共价连接导致的生物活性丧失；（2）我们构建了一个良好封闭的传感器表面，在缓冲液中对每种生物标志物进行单重检测时，非特异性吸附最小；（3）我们采用了夹心分析法，将功能化量子点（纳米增强剂）用作信号放大器，在缓冲液中对KIM-1和HMGB-1实现了5 pg/mL的灵敏度水平。我们预计，将这个多重且超灵敏的平台扩展到评估低浓度和复杂基质中的多种其他新兴临床生物标志物具有巨大潜力并会取得成功。
