Analytical Ultracentrifugation

J. L. Cole, Merck Research Laboratories, West Point, PA, USA

Copyright © 2000 Academic Press

Analytical ultracentrifugation (AUC) involves the measurement of the radial concentration gradients of molecules created by the application of centrifugal force. In contrast to preparative centrifugation, which is used to fractionate mixtures, AUC is a purely analytical technique. Since the pioneering work of Sved-berg and associates in the 1920s, AUC has been employed to characterize the mass, size, shape and association properties of macromolecules in solution. The technique has been broadly applied to research problems in biochemistry, molecular biology and polymer sciences and has also found practical applications in the pharmaceutical and biotechnology industries. Some of the most attractive features of AUC are:

1. Versatility: a wide variety of samples can be examined by AUC, including molecules ranging in size from sucrose to virus particles.

2. Rigor: AUC experiments are directly interpreted in the context of thermodynamic and hy-drodynamic theory, so it is not necessary to run standards to calibrate each experiment.

Also, because the experiments are performed in free solution there are no complications due to interactions with matrices or surfaces that can complicate interpretation of other types of measurements. 3. Convenience: recently, new instrumentation (Beckman Coulter XL-A and XL-I) and data analysis methods have made AUC much more convenient and accessible to the general biochemistry and polymer science communities. In contrast to earlier instruments, experiments are easy to set up and centrifugation parameters and data acquisition are all under computer control. In addition, powerful desktop computers and new software have greatly accelerated the data analysis process and have also extended the capabilities of AUC.

A complete treatment of the theory and applications of AUC is beyond the scope of this article, and the interested reader is referred to the Further Reading section.

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