The Use of Monoclonal Antibodies to Human Platelet Membrane Glycoprotein IIb-IIIa to Quantitate Platelet Deposition on Artificial Surfaces

Juliette N Mulvihill; Han G Huisman; Jean-Pierre Cazenave; Jan A van Mourik; Willem G van Aken

doi:10.1055/s-0038-1645980

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1987; 58(02): 724-731
DOI: 10.1055/s-0038-1645980

Original Articles

Schattauer GmbH Stuttgart

The Use of Monoclonal Antibodies to Human Platelet Membrane Glycoprotein IIb-IIIa to Quantitate Platelet Deposition on Artificial Surfaces

Authors

Juliette N Mulvihill

INSERM U.311, Biologie et Pharmacologie des Interactions du Sang avec les Vaisseaux et les Biomatériaux, Centre Régional de Transfusion Sanguine, Strasbourg, France
Han G Huisman

^*Central Laboratory of The Netherlands Red Cross Blood Transfusion Service and the Laboratory for Experimental and Clinical Immunology, University of Amsterdam, The Netherlands
Jean-Pierre Cazenave

INSERM U.311, Biologie et Pharmacologie des Interactions du Sang avec les Vaisseaux et les Biomatériaux, Centre Régional de Transfusion Sanguine, Strasbourg, France
Jan A van Mourik

^*Central Laboratory of The Netherlands Red Cross Blood Transfusion Service and the Laboratory for Experimental and Clinical Immunology, University of Amsterdam, The Netherlands
Willem G van Aken

^*Central Laboratory of The Netherlands Red Cross Blood Transfusion Service and the Laboratory for Experimental and Clinical Immunology, University of Amsterdam, The Netherlands

Abstract

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Summary

A new technique is described to quantitate platelet deposition in vitro on artifical surfaces, based on a surface phase radioimmunoassay using the monoclonal antibody 6C9, directed specifically against the membrane glycoprotein complex IIb-IIIa of human platelets. Results correlate in linear fashion with those obtained using ¹¹¹Indium labeled platelets. The method offers particular advantages for the measurement of platelet deposition in whole blood, since platelet separation, washing and labeling procedures are eliminated, together with the ensuing possible selection of platelet populations. In vitro perfusion is performed in glass capillaries of precisely defined diameter (0.80 or 0.56 mm i.d.). Blood flow is laminar and accurately controlled over wall shear rates ranging from venous to capillary (50-4,000 s^-1). Using glass capillaries precoated with purified human albumin or fibrinogen or bovine collagen, platelet deposition from suspensions of washed human platelets in Tyrode's-albumin buffer in the presence of a 40% hematocrit is (platelets/mm²): 11,000 (albumin), 78,000 (fibrinogen) and 306,000 (collagen) after 5 min perfusion at 2,000 s^-1. In heparin, citrate or hirudin anticoagulated whole blood, surfaces are passivated, probably by albumin adsorption from plasma (platelets/mm²): 400 (albumin), 3,600 (fibrinogen) and 48,000 (collagen) after 5 min perfusion in the presence of 13 mM citrate.

Keywords

Platelet deposition - Thrombosis - Monoclonal antibody - Glycoprotein IIb-IIIa - Biomaterials - Capillary perfusion

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References

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