SBIR-STTR Award

FLoBio LLC, in conjunction with researchers at the University of Pennsylvania, has pioneered low volume microfluidic assays to monitor blood function under diverse disease and pharmacological conditions. Utilizing over a decade of experience with protein micropatterning, microfluidics, novel fluorescent biosensors, and patient blood testing, the team has developed a point-of-care (POC) disposable chip that allows scalable manufacturing and reliable bedside emergency room (ER) use. Such single-use chips will be developed for rapid patient monitoring in the context of detection of direct oral anticoagulants (DOACs). In over 4 million treatment visits, direct oral anticoagulant (DOAC) such as apixaban, rivaroxaban, or dabigatran are prescribed. DOACs are used for stroke prevention in patients with atrial fibrillation (A-fib) and in the treatment of venous thromboembolism (VTE) or prevention of VTE following hip or knee arthroplasty. These drugs potently inhibit coagulation Factor Xa (FXa) or thrombin (FIIa). Unfortunately, these patients are at greater risk of bleeding following trauma or emergency elective surgery. For example, A-fib patients on DOACs that suffer a stroke are poor candidates for emergency lytic therapy. A technology for monitoring DOAC levels and clearance would address an unmet medical need to identify bleeding risk during trauma or surgery, especially in elder patients with poor renal clearance of a DOAC. FLoBio will implement Phase I research on the following Specific Aims: Manufacture of a single-use chip for detection of FXa inhibitors (FXi) or direct thrombin inhibitors (DTI), especially at levels below <50 ng/mL (< 100 nM) where clinicians specifically need diagnostic information; Development of an alpha unit for chip reading with robust chip operating protocols and data analysis for scoring DOAC levels; and placement of a unit for preliminary on-site tests using blood from consenting, non-emergency A-fib or VTE patients taking DOACs for comparison of chip readout with LCMS gold standard measurements made on stored plasmas. In Phase II, beta-units for chip reading will be developed and larger chip manufacturing runs will be implemented for deployment at a major trauma surgery site in the US and at major thrombosis treatment clinical center. In the US, this technology has the potential to benefit nearly 60,000 patients annually on DOACs that suffer stroke, trauma, or a major bleed.

Public Health Relevance Statement:
NARRATIVE FLoBio LLC proposes to design, manufacture, and validate a novel microfluidic chip to monitor whole blood function including platelet, thrombin, and fibrin production under microfluidic conditions. The development of a technology that is small and easy to use will be essential for bedside monitoring of patients taking direct oral anticoagulants (DOACs) who suffer stroke, trauma, or major bleeding episodes. The need for rapid DOAC monitoring is an urgent unmet need widely recognized by clinicians. In the US, this DOAC detection technology has the potential to benefit nearly 60,000 patients on DOACs that suffer stroke, trauma, or a major bleed.

Project Terms:
Elderly; senior citizen; older person; older adult; later life; late life; geriatric; elders; advanced age; inhibitor/antagonist; inhibitor; Antibodies; Anticoagulants; thrombopoiesis inhibitor; blood thinner; Anticoagulant Drugs; Anticoagulant Agents; Antithrombin III; Heparin Cofactor One; Heparin Cofactor I; Heparin Co-Factor One; Heparin Co-Factor I; Factor Xa Inhibitor; Antithrombin II; Antithrombin 2; Atrial Fibrillation; Auricular Fibrillation; Biological Assay; Biologic Assays; Bioassay; Assay; Biology; Blood; Blood Reticuloendothelial System; Blood coagulation; Blood Clotting; Blood Platelets; Thrombocytes; Platelets; Marrow platelet; Calibration; Liquid Chromatography; Citrates; Color; data interpretation; Data Analysis; Data Analyses; Disorder; Disease; drug/agent; Pharmaceutic Preparations; Medication; Drugs; Pharmaceutical Preparations; Ethylenedinitrilotetraacetic Acid; Ethylenediaminetetraacetic Acid; Edathamil; EDTA; Edetic Acid; Emergencies; Emergency Situation; prothrombase; Thrombokinase; Coagulation Factor Xa; Autoprothrombin C; Activated Factor X; Activated Blood Coagulation Factor X; Factor Xa; Fibrin; Factor One; Factor I; Coagulation Factor One; Coagulation Factor I; Blood Factor One; Blood Coagulation Factor One; Blood Coagulation Factor I; Fibrinogen; flow cytophotometry; Flow Microfluorometry; Flow Microfluorimetry; Flow Cytofluorometry; Flow Cytofluorometries; Flow Cytometry; fluorescent antibody; Immunofluorescence Technique; Immunofluorescence Technic; Fluorescent Antinuclear Antibody Test; Fluorescent Antibody Technic; Coon's Technique; Coon's Technic; Fluorescent Antibody Technique; Gold; Hand; Hematology Testing; Hematological Tests; Hematologic Tests; Blood Tests; hemodynamics; Hemophilia; Factor VIII Deficiency; Hemophilia A; blood loss; Bleeding; Hemorrhage; Hemostasis; Hemostatic function; Modern Man; Human; Methods; optical; Optics; Patents; Legal patent; Patient Monitoring; Patients; Pennsylvania; Pharmacology; Reticuloendothelial System, Serum, Plasma; Plasma Serum; Blood Plasma; Plasma; Printing; Production; Proteins; Scientific Publication; Publications; Reading; Reagent; realtime systems; Real-Time Systems; Research; Researchers; Investigators; Research Personnel; Risk; Running; biological signal transduction; Signaling; Signal Transduction Systems; Intracellular Communication and Signaling; Cell Signaling; Cell Communication and Signaling; Signal Transduction; cerebrovascular accident; cerebral vascular accident; brain attack; Cerebrovascular Stroke; Cerebrovascular Apoplexy; Cerebral Stroke; Brain Vascular Accident; Apoplexy; Stroke; Technology; Testing; fibrinogenase; Thrombase; Thrombin; Thromboembolism; Urothromboplastin; Tissue Thromboplastin; Tissue Factor Procoagulant; Tissue Factor; Prothrombinase; Glomerular Procoagulant Activity; Factor III; Coagulin; Coagulation Factor III; CD142 Antigens; Blood Coagulation Factor III; Thromboplastin; thrombotic disorder; thrombotic disease; Thrombosis; Time; Trypsin Inhibitors; Universities; Viscosity; Measures; total knee arthroplasty; Total Knee Replacement; Knee replacement; Knee joint replacement operation; Knee arthroplasty; knee replacement arthroplasty; Custom; Blood specimen; Blood Sample; Site; Surface; Clinical; Phase; Medical; Link; Renal clearance function; renal clearance; Measurement; Liquid substance; liquid; fluid; Deposition; Deposit; instrument; Venous; Diagnostic; Molds; Filamentous Fungi; Whole Blood; programs; hip replacement arthroplasty; hip replacement; hip joint replacement; hip arthroplasty; Hip Prosthesis Implantation; Investigation; Lytic; Event; Oral; Protocols documentation; Protocol; Route; System; Operative Surgical Procedures; surgery; Surgical Procedure; Surgical Interventions; Surgical; Operative Procedures; interest; Visit; Accident and Emergency department; Emergency room; Emergency Department; experience; mutant; tandem mass spectrometry; Biosensor; biological sensor; novel; Prevention; Devices; Code; Coding System; Sampling; response; Fibrillar Collagen; Molecular Interaction; Binding; µfluidic; Microfluidics; Q-Dot; Quantum Dots; prevent stroke; Stroke prevention; Coagulation; Clotting; Coagulation Process; Address; Microfluidic Microchips; microfluidic chip; Microfluidic Lab-On-A-Chip; Microfluidic Device; Consent; Data; Detection; Reader; Recombinants; Clinical Treatment; trial treatment; trial regimen; Validation; Monitor; transmission process; Transmission; Preparation; Development; developmental; Emergency Care; Emergency medical care; Emergency healthcare; Emergency health care; Emergency Room care; Emergency Department care; ER care; ED care; point of care; Image; imaging; Operating System; preclinical study; pre-clinical study; design; designing; next generation; scale up; Trauma; prototype; manufacturing scale-up; point-of-care diagnostics; operation; trauma care; invention; neonatal surgery; recruit; Injections; off-patent

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.

Public Health Relevance Statement:
Narrative: While direct oral anticoagulants (DOACs) are becoming the preferred approach to managing anticoagulation, DOACs can elevate bleeding risks, and complicate emergency care, which may include the use of expensive DOAC reversal agents. Current diagnostics fail to quickly identify patient DOAC use (type and concentration), and with no FDA approved diagnostic for DOAC detection, 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. FloBio is developing a comprehensive, point of use DOAC test that will provide both DOAC type and relative concentration of DOAC in a patient's blood, within 10 minutes, thereby providing for informed critical care decision making that should reduce both DOAC related complications, and critical care expense.

Project Terms:
Adult; 21+ years old; Adult Human; adulthood; inhibitor; Anticoagulants; Anticoagulant Agents; Anticoagulant Drugs; blood thinner; thrombopoiesis inhibitor; Anticoagulation; Antithrombin III; Antithrombin 2; Antithrombin II; Factor Xa Inhibitor; Heparin Co-Factor I; Heparin Co-Factor One; Heparin Cofactor I; Heparin Cofactor One; Arrhythmia; Cardiac Arrhythmia; Heart Arrhythmias; Automation; Biological Assay; Assay; Bioassay; Biologic Assays; Blood; Blood Reticuloendothelial System; Blood Circulation; Bloodstream; Circulation; Blood coagulation; Blood Clotting; Blood Platelets; Marrow platelet; Platelets; Thrombocytes; Blood Vessels; vascular; Capital; Classification; Systematics; Clinical Research; Clinical Study; Critical Care; Decision Making; Disease; Disorder; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Fibrin; Goals; Gold; Health Personnel; Health Care Providers; Healthcare Providers; Healthcare worker; health care personnel; health care worker; health provider; health workforce; healthcare personnel; medical personnel; treatment provider; hemodynamics; Hemophilia A; Factor VIII Deficiency; Hemophilia; Hemorrhage; Bleeding; blood loss; Human; Modern Man; In Vitro; Investments; Joints; Laboratories; Manuals; Medicine; Patients; Pennsylvania; Physicians; Production; Proteins; Reagent; Risk; Running; Safety; Sensitivity and Specificity; Cell Communication and Signaling; Cell Signaling; Intracellular Communication and Signaling; Signal Transduction Systems; Signaling; biological signal transduction; Signal Transduction; Software; Computer software; Apoplexy; Brain Vascular Accident; Cerebral Stroke; Cerebrovascular Apoplexy; Cerebrovascular Stroke; brain attack; cerebral vascular accident; cerebrovascular accident; Stroke; Testing; Thrombase; fibrinogenase; Thrombin; Blood Coagulation Factor III; CD142 Antigens; Coagulation Factor III; Coagulin; Factor III; Glomerular Procoagulant Activity; Prothrombinase; Tissue Factor; Tissue Factor Procoagulant; Tissue Thromboplastin; Urothromboplastin; Thromboplastin; thrombotic disease; thrombotic disorder; Thrombosis; Time; Collagen Type I; Type 1 Collagen; Universities; Warfarin; Work; Measures; Outcome Assessment; Data Set; Dataset; Blinded; Caring; analytical method; Blood specimen; Blood Sample; improved; Site; Area; Clinical; Phase; Variant; Variation; Physiological; Physiologic; Measurement; Reporter; instrument; Diagnostic; Whole Blood; Knowledge; programs; Event; Oral; Clinic; System; Operative Procedures; Surgical; Surgical Interventions; Surgical Procedure; surgery; Operative Surgical Procedures; meetings; Performance; Devices; Sampling; µfluidic; Microfluidics; Clotting; Coagulation; Coagulation Process; Length; Microfluidic Device; Microfluidic Lab-On-A-Chip; microfluidic chip; Microfluidic Microchips; Data; Detection; Reproducibility; in vivo; Clinical Data; Monitor; Process; Development; developmental; Emergency Care; ED care; ER care; Emergency Department care; Emergency Room care; Emergency healthcare; Emergency medical care; Emergency health care; Image; imaging; novel strategies; new approaches; novel approaches; novel strategy; Outcome; Trauma; innovation; innovate; innovative; evidence base; FDA approved; patient population; product development; point-of-care diagnostics; diagnostic assay; in-vitro diagnostics