Nieuwenhuysen P, Clauwaert J
J Biochem Biophys Methods. 1981 Dec;5(5):279-86. doi: 10.1016/0165-022x(81)90038-5.
It is impossible to measure the diffusion coefficient of macromolecules directly and accurately by quasi-elastic light scattering, when aggregates cannot be eliminated from the solutions to be investigated. Nevertheless, a simple method can be applied to overcome this problem in many cases. Aggregates are separated from the monomeric macromolecules by rate-zonal sedimentation in a CsCl density gradient in a transparent centrifugation tube; the monomers are then located by laser light scattering intensity measurements; photon correlation spectroscopy of the scattered ligh finally yields their diffusion coefficient. The viscosity of aqueous CsCl solutions at different temperatures and concentrations allows a good separation by centrifugation and a low uncertainty in the reduction of the measured diffusion coefficient to standard conditions. The application of the method to eukaryotic large ribosomal subunits is described as an example.
