Macromolecular Size-And-Shape Distributions by Sedimentation Velocity Analytical Ultracentrifugation

¹ National Institutes of Health

^* To whom correspondence should be addressed. E-mail: pschuck{at}helix.nih.gov.

Submitted on January 21, 2006
Revised on February 15, 2006
Accepted on 23 February 2006

	Abstract

Sedimentation velocity analytical ultracentrifugation is an important tool in the characterization of macromolecules and nanoparticles in solution. The sedimentation coefficient distribution c(s) of Lamm equation solutions is based on the approximation of a single, weight-average frictional coefficient of all particles, determined from the experimental data, which scales the diffusion coefficient to the sedimentation coefficient consistent with the traditional s ~ M^2/3 power law. It provides a high hydrodynamic resolution, where diffusional broadening of the sedimentation boundaries is deconvoluted from the sedimentation coefficient distribution. The approximation of a single weight-average frictional ratio is favored by several experimental factors, and usually gives good results for chemically not too dissimilar macromolecules, such as mixtures of folded proteins. In this communication, we examine an extension to a two-dimensional distribution of sedimentation coefficient and frictional ratio, c(s,f_r), which is representative of a more general set of size-and-shape distributions, including mass-Stokes radius distributions, c(M,R_S), and sedimentation coefficient-molar mass distributions c(s,M). We show that this can be used to determine average molar masses of macromolecules and characterize macromolecular distributions, without the approximation of any scaling relationship between hydrodynamic and thermodynamic parameters.

Key Words: Fredholm integral equation, Lamm equation, hydrodynamics, nanoparticles, regularization, size-distribution

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