| Abstract
| - The single biggest problem with solution-phase H/Dexchange as a mass spectrometric probe of surfaceexposure in a protein (or protein complex) is back-exchange of H for D after the initial H/D exchange hasbeen quenched. Back-exchange results in loss of pertinentdata and also greatly hampers data analysis. Previously,very fast, cold (0−4 °C) HPLC was performed to helpreduce back-exchange, but calculated back-exchange stillaverages ∼30%. In this report, supercritical fluid chromatography replaces HPLC as the desalting/separationtechnique prior to mass analysis, providing a dramaticreduction in back-exchange compared to the fast, coldHPLC methods.
|
