Evaluation of Polystryene-Divinylbenzene Cross-Linked Chromatographic Media for Seperation of Proteins
Reversed-phase chromatography of proteins on silica based
columns commonly suffers from broad bands and incomplete recovery.
Sixteen experimental porous polystyrene-divinylbenzene (PS/DVB) media
with various average pore sizes, surface areas, degrees of
cross-linking, and particle sizes were evaluated along with four
commercially available PS/DVB media. Ribonuclease A, bovine serum
albumin, beta-lactoglobulin A, and ovalbumin were used as a model
sample to assess the PS/DVB media. PS/DVB performance was evaluated by
peak capacity, protein recovery, and carryover. Peak capacity was
calculated as the number of peaks separated in a 15 minute gradient.
Carryover was evaluated by three successive blank gradients following
the sample injection. Average peak capacity varied from 90 to 140 and
protein recovery varied between 30 to 100%. These new PS/DVB sorbents
provide good separation, great recovery, and excellent pH stability
(1-14) for a broad range of mobile phases. This chemical stability
facilitates column regeneration with strong base or acid.