Development and validation of a high throughput whole blood thrombolysis plate assay

Abstract The objective of this work was to develop a high throughput assay for testing in vitro the thrombolytic activity using citrated whole blood samples, and to overcome the limitations of currently available techniques. We successfully developed a method that involves forming halo shaped, tissu...

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Autores principales:	T. Bonnard, L. S. Law, Z. Tennant, C. E. Hagemeyer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/37929394349340edaa11c203ea3edf74
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spelling	oai:doaj.org-article:37929394349340edaa11c203ea3edf742021-12-02T11:53:09ZDevelopment and validation of a high throughput whole blood thrombolysis plate assay10.1038/s41598-017-02498-22045-2322https://doaj.org/article/37929394349340edaa11c203ea3edf742017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02498-2https://doaj.org/toc/2045-2322Abstract The objective of this work was to develop a high throughput assay for testing in vitro the thrombolytic activity using citrated whole blood samples, and to overcome the limitations of currently available techniques. We successfully developed a method that involves forming halo shaped, tissue factor induced, whole blood clots in 96 well plates, and then precisely measuring the thrombolysis process with a spectrophotometer plate reader. We here describe the implementation of this novel method, which we refer to as halo assay, and its validation with plasmin, urokinase and tissue plasminogen activator at different doses. The resulting data is a highly detailed thrombolysis profile, allowing comparison of different fibrinolytic agents. The time point analysis allows kinetic data to be collected and calculated to determine key parameters such as the activation time and the rate of fibrinolysis. We also assessed the capacity of the model to study the effect of clot maturation time on the fibrinolytic rate, an aspect of thrombosis rather unexplored with currently available methods, but of increasing importance in drug development. This novel thrombolysis assay could be an extremely useful research tool; to study the complex process of thrombolysis, and a valuable translational clinical tool; as a screening device to rapidly identify hypo- or hyper-fibrinolysis.T. BonnardL. S. LawZ. TennantC. E. HagemeyerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q T. Bonnard L. S. Law Z. Tennant C. E. Hagemeyer Development and validation of a high throughput whole blood thrombolysis plate assay
description	Abstract The objective of this work was to develop a high throughput assay for testing in vitro the thrombolytic activity using citrated whole blood samples, and to overcome the limitations of currently available techniques. We successfully developed a method that involves forming halo shaped, tissue factor induced, whole blood clots in 96 well plates, and then precisely measuring the thrombolysis process with a spectrophotometer plate reader. We here describe the implementation of this novel method, which we refer to as halo assay, and its validation with plasmin, urokinase and tissue plasminogen activator at different doses. The resulting data is a highly detailed thrombolysis profile, allowing comparison of different fibrinolytic agents. The time point analysis allows kinetic data to be collected and calculated to determine key parameters such as the activation time and the rate of fibrinolysis. We also assessed the capacity of the model to study the effect of clot maturation time on the fibrinolytic rate, an aspect of thrombosis rather unexplored with currently available methods, but of increasing importance in drug development. This novel thrombolysis assay could be an extremely useful research tool; to study the complex process of thrombolysis, and a valuable translational clinical tool; as a screening device to rapidly identify hypo- or hyper-fibrinolysis.
format	article
author	T. Bonnard L. S. Law Z. Tennant C. E. Hagemeyer
author_facet	T. Bonnard L. S. Law Z. Tennant C. E. Hagemeyer
author_sort	T. Bonnard
title	Development and validation of a high throughput whole blood thrombolysis plate assay
title_short	Development and validation of a high throughput whole blood thrombolysis plate assay
title_full	Development and validation of a high throughput whole blood thrombolysis plate assay
title_fullStr	Development and validation of a high throughput whole blood thrombolysis plate assay
title_full_unstemmed	Development and validation of a high throughput whole blood thrombolysis plate assay
title_sort	development and validation of a high throughput whole blood thrombolysis plate assay
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/37929394349340edaa11c203ea3edf74
work_keys_str_mv	AT tbonnard developmentandvalidationofahighthroughputwholebloodthrombolysisplateassay AT lslaw developmentandvalidationofahighthroughputwholebloodthrombolysisplateassay AT ztennant developmentandvalidationofahighthroughputwholebloodthrombolysisplateassay AT cehagemeyer developmentandvalidationofahighthroughputwholebloodthrombolysisplateassay
_version_	1718394860125290496

Development and validation of a high throughput whole blood thrombolysis plate assay

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