Von Willebrand factor cleaving protease activity in the physiopathology of microangiopathic disorders
Medicina (B.Aires)
;
63(2): 130-136, 2003. tab, graf
Artículo
en Inglés
| LILACS
| ID: lil-338577
RESUMO
The von Willebrand factor cleaving protease (VWFCP) modulates the von Willebrand factor (VWF) multimeric size in normal plasma. VWFCP activity levels are decreased in different physiological and pathologic situations. Different techniques have been developed to unfold the purified VWF (perfusion at high shear rate, dialysis against urea in nitrocellulose filters), to detect the VWFCP activity on it (multimeric analysis of VWF, collagen binding to VWF assay) and to use the patient plasma both as the source of the enzyme and substrate. In this paper we compared the above mentioned methods with new ones normal plasma dialyzed on membranes instead of purified VWF, dialysis of the samples against urea in tubing instead of nitrocellulose filters, and sonicated plasma to remove the endogenous VWF. The perfusion assay and detection by multimeric analysis showed a limit of detection (25%) of VWFCP activity. Dialysis against urea in both supports and detection by multimeric analysis, showed a better limit of detection (3%), but the recovery of the samples was not as efficient in nitrocellulose filters as it was in tubing. The detection by collagen binding to VWF has more advantages because it allows to analyze more samples than the multimeric analysis does in the same assay. The dialysis of plasma by membranes to obtain the source of exogenous VWF requires no complex equipment. The method, which uses patient plasma as the source of the enzyme and substrate, was inapplicable in our experience because the values could not be interpolated in the reference curve
Texto completo:
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Índice:
LILACS (Américas)
Asunto principal:
Púrpura Trombocitopénica Trombótica
/
Factor de von Willebrand
/
Metaloendopeptidasas
Límite:
Humanos
Idioma:
Inglés
Revista:
Medicina (B.Aires)
Asunto de la revista:
Medicina
Año:
2003
Tipo del documento:
Artículo
País de afiliación:
Argentina
Institución/País de afiliación:
Academia Nacional de Medicina/AR
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