Characterization of BSA unfolding and aggregation using a single-capillary viscometer and dynamic surface tension detector |
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Authors: | Bramanti Emilia Ferrari Carlo Angeli Valeria Onor Massimo Synovec Robert E |
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Institution: | a 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 b 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 c Department of Chemistry, Box 351700, Center for Process Analytical Chemistry, University of Washington, Seattle, WA 98195, USA |
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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 μL 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 3σ of the blank, were 0.012 cP and 0.6 dyn cm−1, respectively.The FI-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. |
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Keywords: | Denaturation β-Aggregation Proteins Viscosity Surface tension DSTD Bovine serum albumin FTIR conformational analysis |
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