Abstract
In 1926, the Swedish scientist Theodor Svedberg was awarded the Nobel Prize in Chemistry for his work on a disperse system, and for studying the colloidal properties of proteins. This work was, to a large extent, made possible by his invention of a revolutionary tool, the analytical ultracentrifuge. These days, technological advances in hardware and computing have transformed the field of analytical ultracentrifugation (AUC) by enabling entirely new classes of experiments and modes of measurement unimaginable by Svedberg, making AUC once again an indispensable tool for modern biomedical research. In this article these advances and their impact on studies of interacting molecules will be discussed, with particular emphasis on a new method termed multi-wavelength analytical ultracentrifugation (MWL-AUC). Novel detectors allow us to add a second dimension to the separation of disperse and heterogeneous systems: in addition to the traditional hydrodynamic separation of colloidal mixtures, it is now possible to identify the sedimenting molecules by their spectral absorbance properties. The potential for this advance is significant for the study of a large range of systems. A further advance has occurred in data management and computational capabilities, opening doors to improved analysis methods, as well as direct networking with the instrument, facilitating data acquisition and data handling, and significant increases in data density from faster detectors with higher resolution capability.
Highlights
analytical ultracentrifugation (AUC) is a biophysical technique that uses centrifugal force to separate molecules dissolved in solution based on their size, anisotropy and buoyant density
The primary variables determined from AUC experiments are the sedimentation, and the diffusion coefficients and the partial concentrations
Zhang, J., Pearson, J.Z., Gorbet, G.E. et al (2017) Spectral and hydrodynamic analysis of West Nile Virus RNA–protein interactions by multiwavelength sedimentation velocity in the analytical ultracentrifuge
Summary
AUC is a biophysical technique that uses centrifugal force to separate molecules dissolved in solution based on their size, anisotropy and buoyant density. The primary variables determined from AUC experiments are the sedimentation, and the diffusion coefficients and the partial concentrations. Biophysics of any molecules present in a mixture of solutes. These parameters can be used to obtain additional information about other molecular properties (see Box 1 for details). The experimental sedimentation and diffusion coefficients are determined by fitting an optical signal trace, obtained over the radial range of the sample compartment at multiple time points (see Figure 1), to finite element solutions of the Lamm equation, a partial differential equation that describes the sedimentation and diffusional transport in an AUC cell. Due to the complexity of the fitting calculation these computations are typically performed on supercomputers to maximize speed and resolution of the analysis
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