title: Individual platelet adhesion assay: measuring platelet function and antiplatelet therapies in whole blood via digital quantification of cell adhesion.
abstract: Widespread monitoring of platelet function and the effect of antiplatelet drugs will improve outcomes in cardiovascular patients, but platelet function testing is not routine in clinical practice. We report a rapid, accurate methodology to quantify platelet-protein interactions: a microarray of contact-printed 6-μm fibrinogen dots on a transparent substrate binds platelets from whole blood, one platelet per dot. The fractional occupancy of an array of fibrinogen dots after a predefined incubation time quantitatively assays platelet adhesion to the protein matrix. We demonstrate this technique by measurement of platelet adhesion to fibrinogen as a means to quantify the effect of the P2Y12 and αIIbβ3 receptor inhibitors cangrelor and abciximab, respectively, both in vitro--by incubating the drug with a freshly drawn blood sample--and in blood from patients treated with antiplatelet agents. The effects of single- and dual-antiplatelet therapy are also assessed. Results from this platelet-binding assay are well correlated with standard techniques including flow cytometry and light transmission aggregometry. This assay technology, readily integrated with microfluidic platforms, is generally applicable to the assay of cell-protein interactions and promises more effective, rapid assay of drug effects in cardiovascular disease patients.
authors.affiliation: Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, Dublin 2, Ireland.
authors.forename: Ana
authors.initials: A
authors.lastname: Lopez-Alonso
authors.forename: Bincy
authors.initials: B
authors.lastname: Jose
authors.forename: Martin
authors.initials: M
authors.lastname: Somers
authors.forename: Karl
authors.initials: K
authors.lastname: Egan
authors.forename: David P
authors.initials: DP
authors.lastname: Foley
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Ricco
authors.forename: Sofia
authors.initials: S
authors.lastname: Ramström
authors.forename: Lourdes
authors.initials: L
authors.lastname: Basabe-Desmonts
authors.forename: Dermot
authors.initials: D
authors.lastname: Kenny
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D006801:Humans; D010973:Platelet Adhesiveness / Q000187:drug effects / Q000502:physiology; D010975:Platelet Aggregation Inhibitors / Q000032:analysis* / Q000494:pharmacology; D010979:Platelet Function Tests / Q000379:methods; D011485:Protein Binding / Q000502:physiology; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D010975:Platelet Aggregation Inhibitors