Effect of protein conformation on experimental bandwidths in reversed-phase high-performance liquid chromatography

K. L. Richards, M. I. Aguilar, M. T.W. Hearn

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Abstract

The dynamic behaviour of a series of closely related cytochrome c molecules has been investigated in reversed-phase high-performance liquid chromatography. In particular, the influence of temperature and gradient time on the experimental bandwidths of equine, tuna, canine and bovine cytochrome c and equine and tuna apocytochrome c has been determined with a C18 and a C4 sorbent. The observed bandbroadening changes are discussed in comparison to the corresponding retention behaviour of the cytochrome c molecules reported in the preceding paper [1]. The results demonstrate that bandwidth measurements can be used as an indicator of the conformational status of a protein under a particular set of chromatographic conditions.

Original languageEnglish
Pages (from-to)33-41
Number of pages9
JournalJournal of Chromatography A
Volume676
Issue number1
DOIs
Publication statusPublished - 29 Jul 1994

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