Protein diffusion coefficient measurements by laminar flow analysis: method and applications

Abstract

Laminar flow in an open tube leads to solute band spreading. This process is opposed by radial diffusion. The translational diffusion coefficient of a solute can be calculated from the band dispersion, tube length, and flow rate. No calibration is necessary. A simple apparatus was used to measure the diffusion coefficients of several small solutes and 18 proteins. Most results were within +/- 10% of literature values. Each determination required 7-90 min, depending on the magnitude of the diffusion coefficient. The amount of protein needed was approximately 25 micrograms. The diffusion coefficient data were useful for the estimation of molecular weights.