Evaluation of Polystryene-Divinylbenzene Cross-Linked Chromatographic Media for Seperation of Proteins

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Peter J. Lee;Amy E. Daly;Martin Gilar;John C. Gebler [Waters]
Fall ACS 2002; 224th ACS National Meeting; Boston August 18-22
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Content Subtype:
Proteins; Ribonuclease A; Bovine serum albumin; beta-lactoglobulin A; Ovalbumin
Symmetry300(™) C4 300 Å 5 µm Steel 4.6 mm 50 mm Delta-Pak C4 300 Å 5 µm Steel 4.6 mm 50 mm
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.

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