A modified clot-based assay to measure negatively charged procoagulant phospholipids

Abstract Growing evidence supports a role for extracellular vesicles (EVs) in haemostasis and thrombosis due to exposure of negatively charged procoagulant phospholipids (PPL). Current commercial PPL-dependent clotting assays use chemically phospholipid depleted plasma to measure PPL activity. The p...

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Autores principales:	Cathrine Ramberg, S. Jamaly, N. Latysheva, L. Wilsgård, T. Sovershaev, O. Snir, J.-B. Hansen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4e92f3e3a24046bab373fe1dea36872c
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spelling	oai:doaj.org-article:4e92f3e3a24046bab373fe1dea36872c2021-12-02T17:15:16ZA modified clot-based assay to measure negatively charged procoagulant phospholipids10.1038/s41598-021-88835-y2045-2322https://doaj.org/article/4e92f3e3a24046bab373fe1dea36872c2021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-88835-yhttps://doaj.org/toc/2045-2322Abstract Growing evidence supports a role for extracellular vesicles (EVs) in haemostasis and thrombosis due to exposure of negatively charged procoagulant phospholipids (PPL). Current commercial PPL-dependent clotting assays use chemically phospholipid depleted plasma to measure PPL activity. The purpose of our study was to modify the PPL assay by substituting the chemically phospholipid depleted plasma with PPL depleted plasma obtained by ultracentrifugation This in order to get readily access to a sensitive and reliable assay to measure PPL activity in human plasma and cell supernatants. The performance of the assay was tested, including the influence of individual coagulation factors and postprandial lipoproteins and compared to a commercial PPL assay (STA-Procoag-PPL). The two PPL assays displayed similar sensitivity to exogenously added standardized phospholipids. The PPL activity measured by the modified assay strongly correlates with the results from the commercial assay. The intraday- and between-days coefficients of variation ranged from 2–4% depending on the PPL activity in the sample. The modified PPL assay was insensitive to postprandial lipoprotein levels in plasma, as well as to tissue factor (TF) positive EVs from stimulated whole blood. Our findings showed that the modified assay performed equal to the comparator, and was insensitive to postprandial lipoproteins and TF+ EVs.Cathrine RambergS. JamalyN. LatyshevaL. WilsgårdT. SovershaevO. SnirJ.-B. HansenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Cathrine Ramberg S. Jamaly N. Latysheva L. Wilsgård T. Sovershaev O. Snir J.-B. Hansen A modified clot-based assay to measure negatively charged procoagulant phospholipids
description	Abstract Growing evidence supports a role for extracellular vesicles (EVs) in haemostasis and thrombosis due to exposure of negatively charged procoagulant phospholipids (PPL). Current commercial PPL-dependent clotting assays use chemically phospholipid depleted plasma to measure PPL activity. The purpose of our study was to modify the PPL assay by substituting the chemically phospholipid depleted plasma with PPL depleted plasma obtained by ultracentrifugation This in order to get readily access to a sensitive and reliable assay to measure PPL activity in human plasma and cell supernatants. The performance of the assay was tested, including the influence of individual coagulation factors and postprandial lipoproteins and compared to a commercial PPL assay (STA-Procoag-PPL). The two PPL assays displayed similar sensitivity to exogenously added standardized phospholipids. The PPL activity measured by the modified assay strongly correlates with the results from the commercial assay. The intraday- and between-days coefficients of variation ranged from 2–4% depending on the PPL activity in the sample. The modified PPL assay was insensitive to postprandial lipoprotein levels in plasma, as well as to tissue factor (TF) positive EVs from stimulated whole blood. Our findings showed that the modified assay performed equal to the comparator, and was insensitive to postprandial lipoproteins and TF+ EVs.
format	article
author	Cathrine Ramberg S. Jamaly N. Latysheva L. Wilsgård T. Sovershaev O. Snir J.-B. Hansen
author_facet	Cathrine Ramberg S. Jamaly N. Latysheva L. Wilsgård T. Sovershaev O. Snir J.-B. Hansen
author_sort	Cathrine Ramberg
title	A modified clot-based assay to measure negatively charged procoagulant phospholipids
title_short	A modified clot-based assay to measure negatively charged procoagulant phospholipids
title_full	A modified clot-based assay to measure negatively charged procoagulant phospholipids
title_fullStr	A modified clot-based assay to measure negatively charged procoagulant phospholipids
title_full_unstemmed	A modified clot-based assay to measure negatively charged procoagulant phospholipids
title_sort	modified clot-based assay to measure negatively charged procoagulant phospholipids
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4e92f3e3a24046bab373fe1dea36872c
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A modified clot-based assay to measure negatively charged procoagulant phospholipids

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