Perfusion chromatography - Characterization of column packings for chromatography of proteins

A Van Deemter equation modeling columns packed with spherical monodisperse perfusive media was utilized to calculate the predicted values of the effective pore diffusivity (D-p) and split ratio (alpha) of columns packed with POROS(R) and Oligo R3(TM) media. The validity of the model was established by calculating consistent values of D-p and alpha for lysozyme, alpha-chymotrypsinogen A and IgG using columns packed with POROS ion-exchange media. The split ratio was shown to be dependent on the pore structure of the media and not the eluite. The technique allows quantitative measurements of alpha and D-p, and is applicable for both perfusive and diffusive media.