Naslov
Supercritical Fluid Chromatography Reduction of Hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis
(Hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis)

Autori
Emmett, Mark R. ; Kazazić, Saša ; Marshall, Alan G. ; Chen, Wei ; Shi, Stone D.-H. ; Bolanos, Ben ; Greig, Michael J.

Izvornik
Analytical Chemistry (0003-2700) 78 (2006), 19; 7058-7060

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
supercritical fluid chromatography; hydrogen/deuterium exchange; mass spectrometry

Sažetak
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 backexchange 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.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

POVEZANOST RADA