Interactions between proteins and ice were studied in situ using FTIR and confocal Raman microspectroscopy under equilibrium and non-equilibrium conditions over a range of temperatures. During quasi-equilibrium freezing of aqueous solutions of dimethyl sulfoxide (DMSO) and bovine serum albumin, preferential exclusion of albumin and/or DMSO was observed. It was hypothesized that the albumin may be adsorbed onto the ice interface or entrapped in the ice phase. To investigate protein-ice interactions during freezing under non-equilibrium conditions, confocal Raman microspectroscopy was used to map the distribution of albumin and the cryoprotective agent trehalose. Microheterogeneity was found in the composition of the freeze-concentrated liquid phase that indicated that albumin was preferentially distributed near or at the boundary of the ice phase. The observed microheterogeneity did not occur under all freezing protocols, which suggests that the technique developed here could be used to develop freezing protocols that would reduce harmful protein-ice interactions.
University of Minnesota Master of Science thesis. April 2013. Major: Mechanical Engineering. Advisor: Alptekin Aksan. 1 computer file (PDF); vii, 93 pages.
Twomey, Alan Michael.
Heterogeneous protein distribution during rapid and equilibrium freezing.
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