Abstract

Introduction: Chromogenic anti-Xa assays are the most appropriate tests to estimate the amount of betrixaban in plasma but the sensitivity of available tests is limited and improvements are needed to encompass the on-therapy range. Methods: Betrixaban was spiked at concentrations ranging from 0 to 500 ng/mL in plasma from healthy donors. Three commercial tests were used (Biophen®DiXaI®, STA®Liquid Anti-Xa, and HemosIL®Liquid Anti-Xa), and adaptation of their sample dilution scheme was performed. These new methodologies were also tested on plasma spiked with amounts of unfractionated heparin (UFH), low molecular weight heparins (LMWH), or fondaparinux covering the on-therapy ranges to evaluate their sensitivity to indirect factor Xa inhibitors. Results: Results showed concentration-dependent decreases in OD/min inversely proportional to the dilutions. While modifications improve the sensitivity of these tests to betrixaban (eg, ½*OD/min of 502 ng/mL [95% CI: 495-508 ng/mL] for Biophen®DiXaI® [1:50] is reduced to 51 ng/mL [95% CI: 50-52 ng/mL] for improved Biophen®DiXaI® [1:5]), results also showed an increased sensitivity to indirect factor Xa inhibitors, except for Biophen®DiXaI® which remains insensitive to UFH and LMWH. Conclusions: Results showed that the improvement of current chromogenic anti-Xa methodologies enhances the sensitivity of these assays to betrixaban but also to indirect factor Xa inhibitors. This lack of specificity may lead to overestimation of betrixaban concentrations in patients bridged with heparins. To avoid this cross-interference, the use of the Biophen®DiXaI® may be a solution except for fondaparinux which remains active even in the presence of the Biophen®DiXaI®’s specific buffer. For the other chromogenic assays, the conception and validation of specific buffer is required.

Original languageEnglish
Number of pages12
JournalInternational Journal of Laboratory Hematology
DOIs
Publication statusPublished - 3 Jan 2019

Fingerprint

Chromogenics
Plasmas
Heparin
Assays
Low Molecular Weight Heparin
Dilution
Buffers
betrixaban
Tissue Donors
Therapeutics
Factor Xa Inhibitors

Keywords

  • betrixaban
  • chromogenic anti-Xa assays
  • factor Xa inhibitors
  • fondaparinux
  • guidance
  • heparins

Cite this

@article{95bac97f13ae40baba3fc843aefd8b70,
title = "Development of new methodologies for the chromogenic estimation of betrixaban concentrations in plasma",
abstract = "Introduction: Chromogenic anti-Xa assays are the most appropriate tests to estimate the amount of betrixaban in plasma but the sensitivity of available tests is limited and improvements are needed to encompass the on-therapy range. Methods: Betrixaban was spiked at concentrations ranging from 0 to 500 ng/mL in plasma from healthy donors. Three commercial tests were used (Biophen{\circledR}DiXaI{\circledR}, STA{\circledR}Liquid Anti-Xa, and HemosIL{\circledR}Liquid Anti-Xa), and adaptation of their sample dilution scheme was performed. These new methodologies were also tested on plasma spiked with amounts of unfractionated heparin (UFH), low molecular weight heparins (LMWH), or fondaparinux covering the on-therapy ranges to evaluate their sensitivity to indirect factor Xa inhibitors. Results: Results showed concentration-dependent decreases in OD/min inversely proportional to the dilutions. While modifications improve the sensitivity of these tests to betrixaban (eg, ½*OD/min of 502 ng/mL [95{\%} CI: 495-508 ng/mL] for Biophen{\circledR}DiXaI{\circledR} [1:50] is reduced to 51 ng/mL [95{\%} CI: 50-52 ng/mL] for improved Biophen{\circledR}DiXaI{\circledR} [1:5]), results also showed an increased sensitivity to indirect factor Xa inhibitors, except for Biophen{\circledR}DiXaI{\circledR} which remains insensitive to UFH and LMWH. Conclusions: Results showed that the improvement of current chromogenic anti-Xa methodologies enhances the sensitivity of these assays to betrixaban but also to indirect factor Xa inhibitors. This lack of specificity may lead to overestimation of betrixaban concentrations in patients bridged with heparins. To avoid this cross-interference, the use of the Biophen{\circledR}DiXaI{\circledR} may be a solution except for fondaparinux which remains active even in the presence of the Biophen{\circledR}DiXaI{\circledR}’s specific buffer. For the other chromogenic assays, the conception and validation of specific buffer is required.",
keywords = "betrixaban, chromogenic anti-Xa assays, factor Xa inhibitors, fondaparinux, guidance, heparins",
author = "Romain Siriez and Jonathan Evrard and Jean-Michel Dogne and Lionel Pochet and C{\'e}line Bouvy and Sarah Lessire and Fran{\cc}ois Mullier and Jonathan Douxfils",
year = "2019",
month = "1",
day = "3",
doi = "10.1111/ijlh.12963",
language = "English",
journal = "International Journal of Laboratory Hematology",
issn = "1751-553X",
publisher = "Wiley-Blackwell Publishing",

}

TY - JOUR

T1 - Development of new methodologies for the chromogenic estimation of betrixaban concentrations in plasma

AU - Siriez, Romain

AU - Evrard, Jonathan

AU - Dogne, Jean-Michel

AU - Pochet, Lionel

AU - Bouvy, Céline

AU - Lessire, Sarah

AU - Mullier, François

AU - Douxfils, Jonathan

PY - 2019/1/3

Y1 - 2019/1/3

N2 - Introduction: Chromogenic anti-Xa assays are the most appropriate tests to estimate the amount of betrixaban in plasma but the sensitivity of available tests is limited and improvements are needed to encompass the on-therapy range. Methods: Betrixaban was spiked at concentrations ranging from 0 to 500 ng/mL in plasma from healthy donors. Three commercial tests were used (Biophen®DiXaI®, STA®Liquid Anti-Xa, and HemosIL®Liquid Anti-Xa), and adaptation of their sample dilution scheme was performed. These new methodologies were also tested on plasma spiked with amounts of unfractionated heparin (UFH), low molecular weight heparins (LMWH), or fondaparinux covering the on-therapy ranges to evaluate their sensitivity to indirect factor Xa inhibitors. Results: Results showed concentration-dependent decreases in OD/min inversely proportional to the dilutions. While modifications improve the sensitivity of these tests to betrixaban (eg, ½*OD/min of 502 ng/mL [95% CI: 495-508 ng/mL] for Biophen®DiXaI® [1:50] is reduced to 51 ng/mL [95% CI: 50-52 ng/mL] for improved Biophen®DiXaI® [1:5]), results also showed an increased sensitivity to indirect factor Xa inhibitors, except for Biophen®DiXaI® which remains insensitive to UFH and LMWH. Conclusions: Results showed that the improvement of current chromogenic anti-Xa methodologies enhances the sensitivity of these assays to betrixaban but also to indirect factor Xa inhibitors. This lack of specificity may lead to overestimation of betrixaban concentrations in patients bridged with heparins. To avoid this cross-interference, the use of the Biophen®DiXaI® may be a solution except for fondaparinux which remains active even in the presence of the Biophen®DiXaI®’s specific buffer. For the other chromogenic assays, the conception and validation of specific buffer is required.

AB - Introduction: Chromogenic anti-Xa assays are the most appropriate tests to estimate the amount of betrixaban in plasma but the sensitivity of available tests is limited and improvements are needed to encompass the on-therapy range. Methods: Betrixaban was spiked at concentrations ranging from 0 to 500 ng/mL in plasma from healthy donors. Three commercial tests were used (Biophen®DiXaI®, STA®Liquid Anti-Xa, and HemosIL®Liquid Anti-Xa), and adaptation of their sample dilution scheme was performed. These new methodologies were also tested on plasma spiked with amounts of unfractionated heparin (UFH), low molecular weight heparins (LMWH), or fondaparinux covering the on-therapy ranges to evaluate their sensitivity to indirect factor Xa inhibitors. Results: Results showed concentration-dependent decreases in OD/min inversely proportional to the dilutions. While modifications improve the sensitivity of these tests to betrixaban (eg, ½*OD/min of 502 ng/mL [95% CI: 495-508 ng/mL] for Biophen®DiXaI® [1:50] is reduced to 51 ng/mL [95% CI: 50-52 ng/mL] for improved Biophen®DiXaI® [1:5]), results also showed an increased sensitivity to indirect factor Xa inhibitors, except for Biophen®DiXaI® which remains insensitive to UFH and LMWH. Conclusions: Results showed that the improvement of current chromogenic anti-Xa methodologies enhances the sensitivity of these assays to betrixaban but also to indirect factor Xa inhibitors. This lack of specificity may lead to overestimation of betrixaban concentrations in patients bridged with heparins. To avoid this cross-interference, the use of the Biophen®DiXaI® may be a solution except for fondaparinux which remains active even in the presence of the Biophen®DiXaI®’s specific buffer. For the other chromogenic assays, the conception and validation of specific buffer is required.

KW - betrixaban

KW - chromogenic anti-Xa assays

KW - factor Xa inhibitors

KW - fondaparinux

KW - guidance

KW - heparins

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U2 - 10.1111/ijlh.12963

DO - 10.1111/ijlh.12963

M3 - Article

JO - International Journal of Laboratory Hematology

JF - International Journal of Laboratory Hematology

SN - 1751-553X

ER -