Protein adsorption onto solid surfaces is an immanent and spontaneous phenomenon that occurs in many fundamental biomedical applications. Recent research mainly focuses on collectively measuring the protein behaviors in confined localized nanosensing areas, whereas ignoring the ambient aqueous variation that directly affects protein behavior, such as the hydrogen bonding network in protein-water which serves as a crucial and mutually constrained environment. Herein, we develop a novel dual-phase plasmonic sensing platform with a gold nanopyramid array (GNPA), simultaneously monitoring protein binding at the nanosensing surface and the variation tendency of hydrogen bonding in aqueous solution. Two types of protein adsorption and their solution variation are explored: we first investigate bovine serum albumin (BSA) adsorption and its variation tendency of hydrogen bonding formation and fracture in water. Then we apply the sensing chip to monitor the fibrinogen adsorption and its real-time conversion to fibrin in an in vitro model of simulated blood coagulation, where the synchronous sensing signals of localized surface plasmon resonance (LSPR) and aqueous hydrogen bonding variations are acquired. It helps to reveal the connection between hydrogen bonding variations in protein-water solution and protein adsorption behaviors at the surface, a meaningful finding for comprehending the initiation of abnormal blood coagulation and the related thromboembolism disease. Moreover, the proposed dual-phase sensing strategy opens a new path for a wider variety of applications, ranging from monitoring microenvironmental changes inside the human body to detecting the water contamination or environmental pollutants.

蛋白质吸附到固体表面是一种内在的自发现象，发生在许多基本的生物医学应用中。最近的研究主要集中在集中测量有限的局部纳米传感区域中的蛋白质行为，而忽略了直接影响蛋白质行为的环境水变化，例如蛋白质-水中的氢键网络，它是一个关键且相互约束的环境。在此，我们开发了一种具有金纳米金字塔阵列（GNPA）的新型双相等离子体传感平台，同时监测纳米传感表面的蛋白质结合和水溶液中氢键的变化趋势。探索了两种类型的蛋白质吸附及其溶液变化：我们首先研究了牛血清白蛋白（BSA）的吸附及其在水中氢键形成和断裂的变化趋势。然后，我们应用传感芯片在体外模拟凝血模型中监测纤维蛋白原吸附及其向纤维蛋白的实时转化，获取局部表面等离子体共振（LSPR）和水氢键变化的同步传感信号。它有助于揭示蛋白质-水溶液中的氢键变化与表面蛋白质吸附行为之间的联系，对于理解异常凝血的开始和相关的血栓栓塞疾病是一个有意义的发现。此外，所提出的双相传感策略为更广泛的应用开辟了一条新途径，