Innovative Biosensing Strategies: Electrochemical Profiling of vWFA2 for Early Sepsis Detection

In the dynamic intersection of technologically advancing medicine and engineering, rapid diagnostic methods are imperative for prognostic evaluation of critically ill - upon multimer assembly patients presenting with clinical complications from a dysregulated host response. Evaluation of biomolecular markers in inflammatory conditions, specifically sepsis, is of peak interest in research to develop a highly rapid, robust, and effective diagnostic option. The complexity of sepsis, a life-threatening inflammatory condition arising from the systemic response to an infectious agent, presents a grave challenge in detection and treatment. Following vascular injury, endothelial cells, strategically placed at the blood-tissue interface, respond to inflammatory stimuli by releasing circulating co-factors, including Von Willebrand Factor (vWF). Von Willebrand Factor-Like 2 Domain (vWFA2), the A2 domain of the heterogenous multimeric glycoprotein, plays a central role in primary hemostasis by promoting platelet adhesion to the subendothelial matrix of damaged vessels and protecting FVIII from proteolytic degradation. vWFA2 responds to shear stress by balancing bleeding and clotting mechanisms. Reduced functionality of vWFA2 leads to severe hemorrhagic consequences following defective formation of a platelet-rich thrombi and fibrin network. The immunethrombotic role of vWFA2 in sepsis is understudied due to the emphasis on its narrowed role in thrombotic events, but it is critically important to understand the developing biological relationship between both physiological processes. Elevated levels of vWF are seen in septic conditions, thus the marker is a vital for disease severity assessment. Current gold-standard diagnostic methods include blood cultures for accurate microbial diagnosis but is faced with compromised sensitivity, prolonged processing time, and a large sample volume requirement. This gap paves the way for development of a prompt and accurate diagnostic protocol, particularly one that targets inflammatory biomarkers. The goal of this research is to develop a biosensor device for rapid detection of biomarkers in septic and inflammatory states. The transformative potential of sensors in revolutionizing detection guides refinement of sensitive assays that addresses the shortcomings of conventional detection methods. This label-free biomolecular assay is fabricated on a flexible hybrid electrode surface and leverages electrochemical impedance spectroscopy (EIS) to measure the capacitive change in impedance, unveiling the binding effects of the target, vWFA2, to the capture probe. The device aims for high sensitivity and specificity in the targeted assay development of a wide dynamic range of 500-32,000 pg/mL. These methods offer a promising solution in point-of-care diagnostic methods, offering a high sensitivity, rapid response time and minimal biofluid volume requirement. The vision behind this research stems from the desire to develop this electrochemical sensor to detect disease states rapidly and accurately, thus creating a pivotal prognostic tool in sepsis treatment, and ultimately mitigating severe mortality and morbidity.