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Characterization of BSA unfolding and aggregation using a single-capillary viscometer and dynamic surface tension
Emilia Bramanti1, Carlo Ferrari, Valeria Angeli
1National 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. bramanti@pi.iccom.cnr.it
Talanta
|October 4, 2011
Summary
A new system combines dynamic surface tension detection (DSTD) with a single capillary viscometer (SCV) for simultaneous measurements. This flow injection analysis method accurately determines viscosity and studies protein denaturation.
Area of Science:
- Analytical Chemistry
- Physical Chemistry
- Biophysical Chemistry
Background:
- Dynamic surface tension detection (DSTD) is a valuable technique for analyzing liquid properties.
- Simultaneous viscosity measurements can provide complementary information, enhancing analytical capabilities.
- Flow injection analysis (FIA) offers rapid and efficient sample processing.
Purpose of the Study:
- To develop and validate a novel system integrating DSTD with a single capillary viscometer (SCV) for simultaneous viscosity and surface tension measurements.
- To assess the performance of the integrated system for flow injection analysis (FIA).
- To apply the system to study complex biological processes like protein denaturation.
Main Methods:
- A dynamic surface tension detector (DSTD) was coupled with a single capillary viscometer (SCV) in a parallel configuration for flow injection analysis.
- Viscosity measurements utilized a PEEK capillary (0.25 mm i.d., 75 cm length) with a differential piezoelectric pressure transducer.
- The system was optimized and validated using water/glycerol mixtures, and applied to analyze bovine serum albumin (BSA) denaturation.
Main Results:
- The integrated SCV-DSTD system demonstrated effective simultaneous measurement of dynamic viscosity and surface tension.
- The SCV achieved a detection limit of 0.012 cP, while the DSTD had a limit of 0.6 dyn cm⁻¹.
- The system successfully monitored temperature-induced denaturation and aggregation of bovine serum albumin (BSA).
Conclusions:
- The combined flow injection analysis system (FI-SCV-DSTD) provides a powerful tool for simultaneous viscosity and surface tension determination.
- The system is capable of characterizing complex biophysical processes, such as protein conformational changes.
- Results obtained were corroborated by Fourier Transform infrared spectroscopy, validating the system's utility.
