Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
45, Number 7, August 2007, pp. 415-421
?Nylon-6 Capillary-Channeled Polymer Fibers as
a Stationary Phase for the Mixed-Mode Ion Exchange/Reversed-Phase
Chromatography Separation of Proteins
?Rayman D. Stanelle, Christine M. Straut, and
R. Kenneth Marcus
Department of Chemistry, Biosystems Research Complex, Clemson
University, Clemson, South Carolina 29634-1905
?Capillary-channeled polymer (C-CP) fibers extruded
from nylon-6 are used as the stationary phase for the ion-exchange/reversed-phase
mixed-mode chromatographic separation of a three protein mixture.
The nylon-6 C-CP fibers are packed collinearly in a 250 ¥ 1.5-mm
i.d. column with an interstitial fraction of ~0.6. The effects
of four displacing salts at three different pHs are studied with
regards to protein retention time, peak width, selectivity, and
resolution for a synthetic mixture consisting of myoglobin, ribonuclease
A, and lysozyme to determine the optimum mobile phase conditions.
The net charge model is found to be inadequate in fully explaining
the retention behavior, as the proteins are retained by anion
and cation-exchange interactions, as well as hydrophobic interactions
with the stationary phase. It is found that pH and displacing
salt strength had a significant influence on the retention properties
and resolution of the proteins.
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