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Updated: Aug 13, 2026

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)
Published on: October 29, 2009
Influence of urea on the high-performance cation-exchange chromatography of hen egg white lysozyme
Abstract:
The effects of urea on the high-performance cation-exchange chromatography of hen egg lysozyme are reported. The capacity factor, k', has been determined as a function of cation concentration with a polyaspartate column using the acetates of Na+, K+, Ca2+ and Mg2+. Urea decreases lysozyme retention. Plots of log k' vs. log ionic strength show linear relationships. The slope of the plot describing the Ca2+ elution of lysozyme was the same in the presence of 5 M urea as in its absence. In strong urea solutions and at elevated temperatures, lysozyme denaturation is evidenced by a marked decrease in k'. The temperature range for denaturation corresponded closely to that observed in intrinsic fluorescence and circular dichroism measurements. The potential utility and limitations of high-performance ion-exchange chromatography for studying protein denaturation are discussed.
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