Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring

Michael Hardy; Julien Cabo; Antoine Deliège; Jonathan Douxfils; Isabelle Gouin-Thibault; Thomas Lecompte; François Mullier

doi:10.1016/j.rpth.2023.102257

Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring

Michael Hardy, Julien Cabo, Antoine Deliège, Jonathan Douxfils, Isabelle Gouin-Thibault, Thomas Lecompte, François Mullier

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Abstract

BACKGROUND: Anti-Xa assays are used for unfractionated heparin (UFH) monitoring. Dextran sulfate (DS) is used in some assays to overcome the artifactual preanalytical release of platelet factor 4. However, the practical implications of this test modification have not been studied extensively.

OBJECTIVES: To investigate the impact of the presence of DS in the anti-Xa assay for UFH laboratory monitoring.

METHODS: We studied factor Xa inhibition, using an assay without DS (Stago Liquid Anti-Xa), in normal pool plasma spiked with various concentrations of UFH (up to 1 IU/mL) in the presence of increasing concentrations of DS (up to 2560 μg/mL). We also investigated the effect of DS on FXa inhibition measured after the addition of UFH and heparin antagonists (protamine and Polybrene; Sigma Aldrich). Eventually, we compared the anti-Xa levels measured using the assay without DS to those measured with an assay containing DS (BIOPHEN Heparin LRT, Hyphen BioMed).

RESULTS: DS per se had a detectable anti-Xa effect. FXa inhibition in UFH-spiked plasma linearly increased with increasing concentrations of added DS, with a plateau at approximately 160 μg/mL DS, at which the apparent anti-Xa level had almost doubled. In the presence of heparin antagonists, the addition of DS increased anti-Xa levels, corresponding to the dissociation of the UFH-antagonists complexes in vitro. With the anti-Xa assay containing DS, UFH inhibition was not detected.

CONCLUSION: In the presence of high concentrations of DS, FXa inhibition was much higher than that predicted from added UFH amounts, presumably related to the greater availability of UFH for interaction with antithrombin. While the relevance of measuring this "masked" heparin has not been demonstrated, the presence of DS renders the result inaccurate in the presence of protamine or Polybrene.

Original language	English
Article number	102257
Pages (from-to)	102257
Journal	Research and Practice in Thrombosis and Haemostasis
Volume	7
Issue number	8
DOIs	https://doi.org/10.1016/j.rpth.2023.102257
Publication status	Published - Nov 2023

Funding

This work was supported by the Fondation Mont-Godinne . This work was supported by the Fondation Mont-Godinne. Conceptualization: M.H. J.C. T.L. F.M. Investigation: M.H. A.D. Formal analysis: M.H. J.C. T.L. F.M. Writing—original draft: M.H. J.C. Writing—review and editing: M.H. J.C. A.D. J.D. I.G.-T. T.L. F.M. Supervision: F.M. All authors have approved the final manuscript. M.H. A.D. J.C. I.G.-T. and T.L. declare no conflict of interest. J.D. is the Chief Executive Officer and founder of QUALIblood s.a. a contract research organization manufacturing the DP-Filter; is a coinventor of the DP-Filter (patent application number: PCT/ET2019/052903); and reports personal fees from Daiichi-Sankyo, Mithra Pharmaceuticals, Stago, Roche, and Roche Diagnostics, outside the submitted work. F.M. reports institutional fees from Stago, Werfen, Nodia, Roche Sysmex, and Bayer and grant or contract from the National Fund for Scientific Research, Belgium. He also reports speaker fees from Boehringer-Ingelheim, Bayer Healthcare, Bristol-Myers Squibb-Pfizer, Stago, Sysmex, and Aspen, all outside the submitted work. M.H., A.D., J.C., I.G.-T., and T.L. declare no conflict of interest. J.D. is the Chief Executive Officer and founder of QUALIblood s.a., a contract research organization manufacturing the DP-Filter; is a coinventor of the DP-Filter (patent application number: PCT/ET2019/052903); and reports personal fees from Daiichi-Sankyo, Mithra Pharmaceuticals, Stago, Roche, and Roche Diagnostics, outside the submitted work. F.M. reports institutional fees from Stago, Werfen, Nodia, Roche Sysmex, and Bayer and grant or contract from the National Fund for Scientific Research, Belgium. He also reports speaker fees from Boehringer-Ingelheim, Bayer Healthcare, Bristol-Myers Squibb-Pfizer, Stago, Sysmex, and Aspen, all outside the submitted work.

Funders	Funder number
Bristol Myers Squibb
Fondation Mont-Godinne	PCT/ET2019/052903
Bayer
Fonds de la Recherche Scientifique F.R.S.-FNRS

Keywords

anti-Xa
dextran
heparin
polybrene
protamine
unfractionated heparin

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.rpth.2023.102257

9861-2023-HardyProof, 1.05 MBLicence: CC BY-NC-ND

Cite this

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title = "Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring",

abstract = "BACKGROUND: Anti-Xa assays are used for unfractionated heparin (UFH) monitoring. Dextran sulfate (DS) is used in some assays to overcome the artifactual preanalytical release of platelet factor 4. However, the practical implications of this test modification have not been studied extensively.OBJECTIVES: To investigate the impact of the presence of DS in the anti-Xa assay for UFH laboratory monitoring.METHODS: We studied factor Xa inhibition, using an assay without DS (Stago Liquid Anti-Xa), in normal pool plasma spiked with various concentrations of UFH (up to 1 IU/mL) in the presence of increasing concentrations of DS (up to 2560 μg/mL). We also investigated the effect of DS on FXa inhibition measured after the addition of UFH and heparin antagonists (protamine and Polybrene; Sigma Aldrich). Eventually, we compared the anti-Xa levels measured using the assay without DS to those measured with an assay containing DS (BIOPHEN Heparin LRT, Hyphen BioMed).RESULTS: DS per se had a detectable anti-Xa effect. FXa inhibition in UFH-spiked plasma linearly increased with increasing concentrations of added DS, with a plateau at approximately 160 μg/mL DS, at which the apparent anti-Xa level had almost doubled. In the presence of heparin antagonists, the addition of DS increased anti-Xa levels, corresponding to the dissociation of the UFH-antagonists complexes in vitro. With the anti-Xa assay containing DS, UFH inhibition was not detected. CONCLUSION: In the presence of high concentrations of DS, FXa inhibition was much higher than that predicted from added UFH amounts, presumably related to the greater availability of UFH for interaction with antithrombin. While the relevance of measuring this {"}masked{"} heparin has not been demonstrated, the presence of DS renders the result inaccurate in the presence of protamine or Polybrene.",

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author = "Michael Hardy and Julien Cabo and Antoine Deli{\`e}ge and Jonathan Douxfils and Isabelle Gouin-Thibault and Thomas Lecompte and Fran{\c c}ois Mullier",

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year = "2023",

month = nov,

doi = "10.1016/j.rpth.2023.102257",

language = "English",

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T1 - Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring

AU - Hardy, Michael

AU - Cabo, Julien

AU - Deliège, Antoine

AU - Douxfils, Jonathan

AU - Gouin-Thibault, Isabelle

AU - Lecompte, Thomas

AU - Mullier, François

PY - 2023/11

Y1 - 2023/11

N2 - BACKGROUND: Anti-Xa assays are used for unfractionated heparin (UFH) monitoring. Dextran sulfate (DS) is used in some assays to overcome the artifactual preanalytical release of platelet factor 4. However, the practical implications of this test modification have not been studied extensively.OBJECTIVES: To investigate the impact of the presence of DS in the anti-Xa assay for UFH laboratory monitoring.METHODS: We studied factor Xa inhibition, using an assay without DS (Stago Liquid Anti-Xa), in normal pool plasma spiked with various concentrations of UFH (up to 1 IU/mL) in the presence of increasing concentrations of DS (up to 2560 μg/mL). We also investigated the effect of DS on FXa inhibition measured after the addition of UFH and heparin antagonists (protamine and Polybrene; Sigma Aldrich). Eventually, we compared the anti-Xa levels measured using the assay without DS to those measured with an assay containing DS (BIOPHEN Heparin LRT, Hyphen BioMed).RESULTS: DS per se had a detectable anti-Xa effect. FXa inhibition in UFH-spiked plasma linearly increased with increasing concentrations of added DS, with a plateau at approximately 160 μg/mL DS, at which the apparent anti-Xa level had almost doubled. In the presence of heparin antagonists, the addition of DS increased anti-Xa levels, corresponding to the dissociation of the UFH-antagonists complexes in vitro. With the anti-Xa assay containing DS, UFH inhibition was not detected. CONCLUSION: In the presence of high concentrations of DS, FXa inhibition was much higher than that predicted from added UFH amounts, presumably related to the greater availability of UFH for interaction with antithrombin. While the relevance of measuring this "masked" heparin has not been demonstrated, the presence of DS renders the result inaccurate in the presence of protamine or Polybrene.

AB - BACKGROUND: Anti-Xa assays are used for unfractionated heparin (UFH) monitoring. Dextran sulfate (DS) is used in some assays to overcome the artifactual preanalytical release of platelet factor 4. However, the practical implications of this test modification have not been studied extensively.OBJECTIVES: To investigate the impact of the presence of DS in the anti-Xa assay for UFH laboratory monitoring.METHODS: We studied factor Xa inhibition, using an assay without DS (Stago Liquid Anti-Xa), in normal pool plasma spiked with various concentrations of UFH (up to 1 IU/mL) in the presence of increasing concentrations of DS (up to 2560 μg/mL). We also investigated the effect of DS on FXa inhibition measured after the addition of UFH and heparin antagonists (protamine and Polybrene; Sigma Aldrich). Eventually, we compared the anti-Xa levels measured using the assay without DS to those measured with an assay containing DS (BIOPHEN Heparin LRT, Hyphen BioMed).RESULTS: DS per se had a detectable anti-Xa effect. FXa inhibition in UFH-spiked plasma linearly increased with increasing concentrations of added DS, with a plateau at approximately 160 μg/mL DS, at which the apparent anti-Xa level had almost doubled. In the presence of heparin antagonists, the addition of DS increased anti-Xa levels, corresponding to the dissociation of the UFH-antagonists complexes in vitro. With the anti-Xa assay containing DS, UFH inhibition was not detected. CONCLUSION: In the presence of high concentrations of DS, FXa inhibition was much higher than that predicted from added UFH amounts, presumably related to the greater availability of UFH for interaction with antithrombin. While the relevance of measuring this "masked" heparin has not been demonstrated, the presence of DS renders the result inaccurate in the presence of protamine or Polybrene.

KW - anti-Xa

KW - dextran

KW - heparin

KW - polybrene

KW - protamine

KW - unfractionated heparin

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U2 - 10.1016/j.rpth.2023.102257

DO - 10.1016/j.rpth.2023.102257

M3 - Article

C2 - 38193053

SN - 2475-0379

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JO - Research and Practice in Thrombosis and Haemostasis

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Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring

Abstract

Funding

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Clinical Pharmacology Research Group

The BIOCOAG project : Biological assessment of the control of anticoagulation

Cite this

Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring

Abstract

Funding

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Projects

Clinical Pharmacology Research Group

The BIOCOAG project : Biological assessment of the control of anticoagulation

Cite this