Abstract
This study illustrates the applicability of dynamic light scattering (DLS)-based optical microrheology in generating new insights into the rheological response of dilute protein solutions as they start to form insoluble aggregates under the influence of a thermal stress. The technique is also shown to provide a quick method for measuring the viscosity in protein solutions. The optical microrheological technique, which is based on DLS with improved single scattering detection, is shown here to capture the rich dynamics in these systems, where traditional mechanical rheometry cannot be effectively employed due to low torque generation and high sample volume requirements and the more widely known diffusing wave spectroscopy microrheology technique is not desirable due to the required high probe particle concentrations The study illustrates the careful consideration which must be given to the tracer particle surface chemistry, tracer particle concentration and tracer particle size in order to extract out rheological responses that are truly representative of the underlying protein dynamics and microstructure. We outline a procedure for ensuring that the pitfalls inherent to this type of measurement are avoided.
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Notes
Note, however, that if the scattering of the particles being measured is not significantly larger than the background scattering from the dispersant, then the intercept will be low, even if only single scattering is present. In the microrheology experiments, the scattering from the probes is always expected to be high by comparison, so the condition given above will hold.
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Acknowledgements
We would like to thank Stephen R. Martin (Division of Physical Biochemistry, National Institute for Medical Research, London, UK) for the CD data. We would also like to thank Kyle Williams from Malvern Instruments for the DSV measurements and Mark Pothecary from Malvern Instruments for his help with the SEC measurements.
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Amin, S., Rega, C.A. & Jankevics, H. Detection of viscoelasticity in aggregating dilute protein solutions through dynamic light scattering-based optical microrheology. Rheol Acta 51, 329–342 (2012). https://doi.org/10.1007/s00397-011-0606-6
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DOI: https://doi.org/10.1007/s00397-011-0606-6