Two novel anti-von Willebrand factor monoclonal antibodies.
Von Willebrand Factor is a multimer produced by endothelial cells and megakaryocytes, being stored in intracellular organelles, such as the Weibel-Palade bodies and alpha-granules in endothelial cells and platelets, respectively. This molecule acts as a carrier protein for factor VIIIc, involved in the intrinsic pathway of blood coagulation maintaining its stability in circulation. Von Willebrand Factor also plays an important role in platelet aggregation and adhesion to injured vessel wall. It interacts with platelets through two distinct glycoproteins, GPIb and GPIIb/IIIa. We raised two monoclonal antibodies, ECA-3 and ECA-4, against human umbilical vascular endothelial cells that recognize and immunoprecipitate von Willebrand Factor. Interestingly, ECA-4 monoclonal antibody is able to completely inhibit platelet agglutination induced by ristocetin, suggesting that it binds to von Willebrand Factor close to platelet GPIb binding site. The use of monoclonal antibodies to identify von Willebrand Factor binding regions to factor VIII or platelets has been reported by others. In pulmonary hypertension, abnormalities have been detected on the multimeric structure of the molecule as well as on its proteolytic fragments, by using monoclonal antibodies. Moreover, monoclonal antibodies raised against specific regions of von Willebrand Factor molecule may allow studies of functional abnormalities of this protein in inherited and acquired disorders like subtypes of von Willebrand's disease.
Spadafora-Ferreira
M
M
Heart Institute-InCor, University of São Paulo, School of Medicine, Brazil.
Lopes
A A
AA
Coelho
V
V
Guilherme
L
L
Kalil
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000494:pharmacology; D000915:Antibody Affinity; D018929:Cell Culture Techniques; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D010974:Platelet Aggregation / Q000187:drug effects; D011233:Precipitin Tests; D012310:Ristocetin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014471:Umbilical Veins / Q000737:chemistry / Q000166:cytology; D014841:von Willebrand Factor / Q000276:immunology*
D000911:Antibodies, Monoclonal; D014841:von Willebrand Factor; D012310:Ristocetin