Abstract
The single biggest problem with solution-phase H/D exchange as a mass spectrometric probe of surface exposure in a protein (or protein complex) is back-exchange of H for D after the initial H/D exchange has been quenched. Back-exchange results in loss of pertinent data and also greatly hampers data analysis. Previously, very fast, cold (0-4 °C) HPLC was performed to help reduce back-exchange, but calculated back-exchange still averages ∼30%. In this report, supercritical fluid chromatography replaces HPLC as the desalting/separation technique prior to mass analysis, providing a dramatic reduction in back-exchange compared to the fast, cold HPLC methods.
Original language | English (US) |
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Pages (from-to) | 7058-7060 |
Number of pages | 3 |
Journal | Analytical Chemistry |
Volume | 78 |
Issue number | 19 |
DOIs | |
State | Published - Oct 1 2006 |
Externally published | Yes |
ASJC Scopus subject areas
- Analytical Chemistry