PERFUSION CHROMATOGRAPHY - CHARACTERIZATION OF COLUMN PACKINGS FOR CHROMATOGRAPHY OF PROTEINS

A Van Deemter equation modeling columns packed with spherical monodisp erse perfusive media was utilized to calculate the predicted values of the effective pore diffusivity (D-p) and split ratio (alpha) of colum ns packed with POROS(R) and Oligo R3(TM) media. The validity of the mo del was established by calculating consistent values of D-p and alpha for lysozyme, alpha-chymotrypsinogen A and IgG using columns packed wi th 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 f or both perfusive and diffusive media.