Characterization of BSA unfolding and aggregation using a single-capillary viscometer and dynamic surface tension detector

Year: 2011

Authors: Bramanti E., Ferrari C., Angeli V., Onor M., Synovec R.E.

Autors Affiliation: National Research Council of Italy, C.N.R., Istituto di Chimica dei Composti Organo Metallici-ICCOM-UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa, Italy;
National Research Council of Italy, C.N.R., Istituto Nazionale di Ottica, INO – UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa, Italy;
Department of Chemistry, Box 351700, Center for Process Analytical Chemistry, University of Washington, Seattle, WA 98195, USA

Abstract: A dynamic surface tension detector (DSTD) has been equipped with an additional pressure sensor for simultaneous viscosity measurements, as a detector for flow injection analysis. The viscosity measurement is based on a single capillary viscometer (SCV) placed in parallel configuration with the DSTD. The viscometer in the optimized conditions consists of a PEEK capillary (i.d.=0.25 mm, L=75 cm) kept at constant temperature using a thermostatic bath, which leads on the two sides to the two arms of a differential piezoelectric pressure transducer with a range of 0-35 psi. The DSTD, described previously, measures the changing pressure across the liquid/air interface of 2 mu L. drops repeatedly forming at the end of a capillary. SCV performance has been evaluated by measuring dynamic viscosity of water/glycerol mixtures analysed in flow injection and comparing the results with the values reported in the literature. The detection limits of SCV and DSTD, calculated as 3 sigma of the blank, were 0.012 cP and 0.6 dyn cm(-1), respectively. The Fl-SCV-DSTD system has been applied to the study of temperature-induced denaturation/aggregation process in bovine serum albumin (BSA). The results have been supported and discussed with respect to BSA conformational analysis performed using Fourier Transform infrared spectroscopy. (C) 2011 Elsevier B.V. All rights reserved

Journal/Review: TALANTA (OXF.)

Volume: 85 (5)      Pages from: 2553  to: 2561

More Information: The authors would like to acknowledge Dr Elpidio Tombari for his helpful discussion in setting up SCV detector. This work has been financially supported by PRIN 2008 funding from Italian Ministry of University and Research (Project No. 2008XXAMZT).
KeyWords: Denaturation; beta-Aggregation; Proteins; Viscosity; Surface tension
DOI: 10.1016/j.talanta.2011.08.009

Citations: 16
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