title: A simple enzyme-immunoassay test for von Willebrand factor binding in human arterial subendothelium.
abstract: In order to determine the binding of vWF, subendothelium from everted human umbilical arteries was perfused with dialysed serum containing different concentrations of purified vWF using an annular perfusion chamber at a wall shear rate of 1100 sec-1 for 30 min. After perfusion, control (not perfused) and perfused vessel segments were washed and incubated with a diluted rabbit antibody against human vWF. Then the nonbound anti-vWF from both samples were used to determine indirectly vWF by EIA. Although in our experiments normal vWF serum concentrations were not enough to exert vWF binding, a substantial binding could be attained with vWF levels around 2.5 U/ml. To estimate the pre-existing subendothelial vWF amount, three different experiments were developed: a) diluted IgG from a nonimmunized rabbit, b) a diluted rabbit antibody to human vWF, c) PBS-BSA. After washing, vessel segments were incubated with rabbit antibody to human vWF. After incubation, the nonbound anti-vWF was used to determine indirectly vWF by EIA. The results obtained showed that the amount of pre-existing vWF was approximately 1.1x10(-3) U vWF/cm2 subendothelium.
authors: Instituto de Investigaciones Hematológicas Mariano R. Castex, Academia Nacional de Medicina, Buenos Aires, Argentina.
authors: A C
authors: AC
authors: Kempfer
authors: J P
authors: JP
authors: Frontroth
authors: C
authors: C
authors: Farías
authors: E
authors: E
authors: Bermejo
authors: M A
authors: MA
authors: Lazzari
mesh_terms: D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D010477:Perfusion; D011485:Protein Binding; D014469:Umbilical Arteries / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D014841:von Willebrand Factor