A platelet activation-specific monoclonal antibody that recognizes a receptor-induced binding site on canine fibrinogen.
An activation-specific monoclonal antibody (MoAb) termed "Canine Activated Platelet 1" (CAP1) has been developed and partially characterized. Flow cytometric studies of isolated canine platelets, using adenosine diphosphate (ADP) and platelet activating factor (PAF) as agonists, demonstrated that CAPI binding site number was proportional to agonist strength and agonist concentration. MoAb CAP1 binding was diminished by ethylenediamine-tetraacetic acid, suggesting that the antigen was either stabilized by calcium or antigen binding to the platelet surface was mediated by calcium. ADP-activated gel-filtered platelets also demonstrated reduced binding of MoAB CAP1 even in the presence of 1 mM CaCl2. Binding of MoAb CAP1 could be partially restored by activating gel-filtered platelets with PAF, suggesting that the antigen was either present within platelet granule membranes or was exposed after binding of released proteins(s) with a platelet receptor. A monoclonal antibody to human platelet glycoprotein IIIa (GPIIIa), which cross-reacts with canine platelet GPIIIa regardless of platelet activation status, did not inhibit binding of MoAb CAP1. MoAb CAP1 bound to isolated canine fibrinogen captured on polystyrene microtiter plates in the absence of platelet proteins. Immunoblots indicated that MoAb CAP1 recognizes nonreduced fibrinogen as well as a plasmin digest of isolated canine fibrinogen. Results of the present studies suggest that MoAb CAP1 recognizes a receptor-induced binding site on canine fibrinogen.
Boudreaux
M K
MK
Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL.
Panangala
V S
VS
Bourne
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism*; D001666:Binding Sites, Antibody / Q000187:drug effects* / Q000276:immunology*; D001667:Binding, Competitive / Q000276:immunology; D004285:Dogs; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism*; D005340:Fibrinogen / Q000276:immunology* / Q000378:metabolism*; D005434:Flow Cytometry; D015539:Platelet Activation / Q000276:immunology*; D010980:Platelet Membrane Glycoproteins / Q000276:immunology* / Q000494:pharmacology*; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor; D005340:Fibrinogen