Measurement of GPV released by activated platelets using a sensitive immunocapture ELISA - Its use to follow platelet storage in transfusion

Citation
Do. Azorsa et al., Measurement of GPV released by activated platelets using a sensitive immunocapture ELISA - Its use to follow platelet storage in transfusion, THROMB HAEM, 81(1), 1999, pp. 131-138
Citations number
31
Categorie Soggetti
Cardiovascular & Hematology Research
Journal title
THROMBOSIS AND HAEMOSTASIS
ISSN journal
03406245 → ACNP
Volume
81
Issue
1
Year of publication
1999
Pages
131 - 138
Database
ISI
SICI code
0340-6245(199901)81:1<131:MOGRBA>2.0.ZU;2-C
Abstract
Thrombin. the most potent platelet agonist, plays a central role in haemost asis and in the occurrence of thrombotic events. This agonist activates pla telets by cleaving the PAR G-protein coupled receptors and by binding to gl ycoprotein (GP) Ib and also cleaves GPV at the platelet surface to liberate the soluble 69 kDa fragment GPVf1. Monoclonal antibodies (MoAbs) to GPV we re developed as tools to study the mechanism of platelet GPV cleavage and m easure release of GPV in pathological situations. Specificity of the MoAbs for GPV was confirmed by flow cytometry and immunoprecipitation of proteins from human platelets and Dami megakaryocytic cells. A sensitive immunocapt ure sandwich ELISA for soluble GPV was developed using two MoAbs recognizin g different epitopes of GPV and purified platelet or recombinant GPV as ref erence protein. This ELISA was employed to determine the mean plasma concen tration of GPV in 100 normal individuals (17.3 ng/ml), to demonstrate the d ose-dependent release of GPVf1 from washed platelets stimulated with thromb in and to follow the progressive release of GPVf1 during storage of therape utic platelet concentrates. The present report describes a sensitive GPV EL ISA of direct application to survey the processing and storage of platelet concentrates for transfusion and of potential value to monitor platelet act ivation in thrombotic states.