BACKGROUND: B-type natriuretic peptide (BNP) is a cardiac hormone that regulates hemodynamic equilibrium. In the circulation, its activity is controlled by proteolytic factors. Accurate measurement of BNP in a patient's plasma may be affected by degradation due to proteolysis. OBJECTIVE: We report on the identification and performance of classes of protease inhibitors that stabilize BNP in plasma. DESIGN AND METHODS: Using the Bayer ADVIA Centaur BNP assay, we measured the effect of arginine, serine and/or specific kallikrein protease inhibitors (PIs) on exogenous spiked or endogenous BNP in patient plasma. RESULTS: Compared to controls without inhibitor, all PIs were capable, to varying degrees, of retarding the rate of proteolytic degradation. The kallikrein-specific inhibitor, D-Phe-Phe-Arg-chloromethylketone (PPACK II) was most effective as a single constituent and was able to eliminate BNP degradation in patient samples for up to 6-10 days when stored at 2-8 degrees C. CONCLUSIONS: The stability of BNP was markedly increased in the presence of kallikrein-specific PPACK II and a broad spectrum of serine PIs. Use of these compounds offers a simple method of extending sample handling and storage of plasma samples containing BNP.
BACKGROUND:B-type natriuretic peptide (BNP) is a cardiac hormone that regulates hemodynamic equilibrium. In the circulation, its activity is controlled by proteolytic factors. Accurate measurement of BNP in a patient's plasma may be affected by degradation due to proteolysis. OBJECTIVE: We report on the identification and performance of classes of protease inhibitors that stabilize BNP in plasma. DESIGN AND METHODS: Using the Bayer ADVIA Centaur BNP assay, we measured the effect of arginine, serine and/or specific kallikrein protease inhibitors (PIs) on exogenous spiked or endogenous BNP in patient plasma. RESULTS: Compared to controls without inhibitor, all PIs were capable, to varying degrees, of retarding the rate of proteolytic degradation. The kallikrein-specific inhibitor, D-Phe-Phe-Arg-chloromethylketone (PPACK II) was most effective as a single constituent and was able to eliminate BNP degradation in patient samples for up to 6-10 days when stored at 2-8 degrees C. CONCLUSIONS: The stability of BNP was markedly increased in the presence of kallikrein-specific PPACK II and a broad spectrum of serine PIs. Use of these compounds offers a simple method of extending sample handling and storage of plasma samples containing BNP.
Authors: Roberto Ferrari; J Cardoso; M C Fonseca; C Aguiar; J I Moreira; A Fucili; C Rapezzi Journal: Clin Res Cardiol Date: 2019-09-17 Impact factor: 5.460
Authors: Laura García-Otero; Marta López; Mariona Guitart-Mampel; Constanza Morén; Anna Goncé; Carol Esteve; Laura Salazar; Olga Gómez; Josep María Martínez; Berta Torres; Sergi César; Glòria Garrabou; Fàtima Crispi; Eduard Gratacós Journal: PLoS One Date: 2019-03-04 Impact factor: 3.240
Authors: Mariona Guitart-Mampel; Diana L Juarez-Flores; Lina Youssef; Constanza Moren; Laura Garcia-Otero; Vicente Roca-Agujetas; Marc Catalan-Garcia; Ingrid Gonzalez-Casacuberta; Ester Tobias; José C Milisenda; Josep M Grau; Fàtima Crispi; Eduard Gratacos; Francesc Cardellach; Glòria Garrabou Journal: J Cell Mol Med Date: 2019-04-02 Impact factor: 5.310