Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis

Mark Emmett, Sasa Kazazic, Alan G. Marshall, Wei Chen, Stone D H Shi, Ben Bolaños, Michael J. Greig

Research output: Contribution to journalArticle

24 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)7058-7060
Number of pages3
JournalAnalytical Chemistry
Volume78
Issue number19
DOIs
StatePublished - Oct 1 2006
Externally publishedYes

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Deuterium
Hydrogen
Ion exchange
Salt removal
Proteins
Supercritical Fluid Chromatography

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis. / Emmett, Mark; Kazazic, Sasa; Marshall, Alan G.; Chen, Wei; Shi, Stone D H; Bolaños, Ben; Greig, Michael J.

In: Analytical Chemistry, Vol. 78, No. 19, 01.10.2006, p. 7058-7060.

Research output: Contribution to journalArticle

Emmett, Mark ; Kazazic, Sasa ; Marshall, Alan G. ; Chen, Wei ; Shi, Stone D H ; Bolaños, Ben ; Greig, Michael J. / Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis. In: Analytical Chemistry. 2006 ; Vol. 78, No. 19. pp. 7058-7060.
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