A rapid microfluidic diagnostic assay for the measurement of Direct Oral Anticoagulants from patient whole blood

Abstract: Direct oral anticoagulants (DOACs) are becoming the preferred approach to managing anticoagulation, particularly in thrombolytic disorders. DOAC use is anticipated to reach $30B globally in the next five years, overtaking other anticoagulants such as warfarin. While highly effective, DOACs can elevate bleeding risks, and complicate emergency care, which may include the use of expensive DOAC reversal agents. As current diagnostics fail to adequately identify problems caused by DOACs, there exists growing safety concerns for this class of drugs that are reinforced by the Joint Commission’s call for laboratory test monitoring, and evidence- based approaches for safe DOAC, and reversal agent use. Whether treating a stroke/VTE/PE patient, heart arrhythmia patient, or trauma/urgent surgery patient, the real-time determination of a patient’s state of anticoagulation, including DOAC type (factor Xa inhibitor vs. direct thrombin inhibitor) is critical for rapid care decisions to avoid adverse bleeding. A significant limitation of core laboratory detection of DOACs is the length of time to results, which can exceed more than the typical 20-minute decision making window. FloBio is developing the first DOAC in vitro diagnostic product to provide physicians with rapid knowledge of anticoagulation state in emergency care settings. With our analyzer/imaging station, and disposable DOAC assay cartridge/microfluidic device, our novel approach combines hemodynamic flow, and discrete clot activation, to mimic physiological blood clotting. In less than 10 minutes, a healthcare provider will know a patient’s relative DOAC concentration, and the DOAC classification. FloBio’s Phase I work has demonstrated a scalable, multiplex, disposable DOAC assay cartridge/microfluidic device that facilitates the natural blood clotting cascade in vitro, an instrument for measuring assay signals, analytic software to provide the end user with a simple outcomes assessment, and clinical data that validates the feasibility of the assay and device to detect DOAC class and level in human blood. FloBio’s innovation is a diagnostic assay that will be the first in the market to provide real-time data on fibrin and platelet accumulation on basal vascular and coagulation proteins, under flow, that recapitulates in vivo coagulation processes. In Phase II, FloBio will focus on validating the DOAC assay in a 100 subject clinical study at the University of Pennsylvania. This study will validate the capability of the assay to detect both class and DOAC level, within a clinical patient population. A second clinical study with DOAC spiked healthy adult blood will determine the device, assay and system range, sensitivity, specificity, variance, and consistency. Clinical assay data will be validated against the gold standard of detection, LCMS. Automation of critical assay functions will also be evaluated to reduce variation in the assay, and the feasibility of a fully integrated, automatable, disposable DOAC assay cartridge will be demonstrated in healthy human blood samples. The outcome of the work will be a fully vetted DOAC assay system that is ready for full development in Phase IIb.