Application of Ultrasonic Shear Rheometer to Characterize Rheological Properties of High Protein Concentration Solutions at Microliter Volume

The purpose of this work was to conduct preliminary rheological analysis on high protein concentration solutions by using the technique of ultrasonic shear rheometry at megahertz frequencies. The work was aimed at establishing the viability of the technique for analyzing protein solution rheology as well as obtaining an initial understanding of the effect of solution conditions on solution rheology of a model protein. Bovine serum albumin (BSA) was used for this study, and rheological analysis was conducted at 20 μL sample volume between pH 2.0 and 9.0 at different ionic strengths at 25°C using 5 and 10 MHz quartz crystals. Significant differences in storage modulus among solutions at pH 5.0, 7.0, and 9.0 could only be detected at 10 MHz, and the errors associated with measurements were smaller as compared to those at 5 MHz for all the solutions studied. Solutions at pH 2.0 and 3.0 showed a time-dependent change in solution rheology. For solutions at pH 5.0, 7.0, and 9.0, which did not show time dependence in solution rheology, loss modulus data at lower concentrations correlated well with the dilute solution data in the literature. At higher concentrations, pH 5.0 solutions exhibited a higher loss modulus than pH 7.0 and pH 9.0 solutions. Storage modulus decreased with increasing ionic strength, unlike loss modulus, which did not show any change, except at pI of protein when no effect was observed. The results show the potential of high frequency rheometry for analyzing subtle differences in rheology of pharmaceutically relevant protein solutions at microliter volume. © 2005 Wiley-Liss, Inc. and the American Pharmacists Association