Rapid Determination of Xa Inhibitor Activity in Blood Using a Microfluidic Device that Measures Platelet Deposition and Fibrin Generation Under Flow

Jason M. Rossi; Karen A. Panckeri; Soumita Ghosh; Tilo Grosser; Adam Cuker; Scott L. Diamond

doi:10.1055/a-2547-5710

TH Open, Inhaltsverzeichnis

CC BY 4.0 · TH Open 2025; 09: a25475710
DOI: 10.1055/a-2547-5710

Original Article

Rapid Determination of Xa Inhibitor Activity in Blood Using a Microfluidic Device that Measures Platelet Deposition and Fibrin Generation Under Flow

Jason M. Rossi

¹Department of Chemical and Biomolecular Engineering, Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania, United States

²FloBio LLC, Philadelphia, Pennsylvania, United States

,

Karen A. Panckeri

³Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States

,

Soumita Ghosh

⁴Institute for Translational Medicine and Therapeutics University of Pennsylvania, Philadelphia, Pennsylvania, United States

,

Tilo Grosser

⁴Institute for Translational Medicine and Therapeutics University of Pennsylvania, Philadelphia, Pennsylvania, United States

⁵Department of Translational Pharmacology, Bielefeld University, Bielefeld, Germany

,

Adam Cuker

³Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States

⁶Department of Pathology & Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States

,

Scott L. Diamond

¹Department of Chemical and Biomolecular Engineering, Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania, United States

⁴Institute for Translational Medicine and Therapeutics University of Pennsylvania, Philadelphia, Pennsylvania, United States

› Institutsangaben

Abstract

Background

Patients taking direct oral anticoagulants (DOACs) often present complicated scenarios following major bleeding, stroke, or emergency surgery. Rapid whole blood assays of DOAC levels would aid clinical decisions such as the need for DOAC reversal.

Methods

We developed a single-use, storage-stable, eight-channel microfluidic device to estimate factor Xa (FXa) inhibitor (apixaban or rivaroxaban) levels in venous thromboembolism or atrial fibrillation patients. The assay simultaneously measured whole blood clotting dynamics on collagen/tissue factor (TF; wall shear rate, 200⁻¹) under four ex vivo conditions: no-treatment control, high dose Factor Xa inhibition, low dose or high dose FXa reversal agent (andexanet alfa). Fibrin and platelet deposition dynamics were monitored via two-color epifluorescence microscopy. Plasma samples were also evaluated by LC-MS/MS for DOAC concentrations.

Results

Experiments with healthy volunteer blood spiked with DOAC verified device performance (DOAC IC₅₀ ∼120 nM) and confirmed that andexanet alfa added to healthy donor blood had no off-target effect on platelet or fibrin signal. Patient whole blood monitored for 15 to 25 minutes (17 minutes mean runtime) allowed calculation of functional DOAC concentrations ranging from 2 to 500 nM that correlated well with LC-MS/MS determination of apixaban or rivaroxaban (R² = 0.7 or 0.9, respectively). Platelet dysfunction was not observed in any patient on DOAC. For a threshold of 100 nM DOAC, the area under the curve (AUC) was found to be 0.881 for apixaban and 0.933 for rivaroxaban.

Conclusion

Microfluidic testing of whole blood can provide a rapid estimate of DOAC levels over the on-therapy range.

Keywords

oral anticoagulant - fibrin - thrombin - microfluidic

Bibliographical Record
Jason M. Rossi, Karen A. Panckeri, Soumita Ghosh, Tilo Grosser, Adam Cuker, Scott L. Diamond. Rapid Determination of Xa Inhibitor Activity in Blood Using a Microfluidic Device that Measures Platelet Deposition and Fibrin Generation Under Flow. TH Open 2025; 09: a25475710.
DOI: 10.1055/a-2547-5710

Volltext

Referenzen

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