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

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

doi:10.1021/ac060693n

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

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

Research output: Contribution to journal › Article

24 Scopus citations

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)
Pages (from-to)	7058-7060
Number of pages	3
Journal	Analytical Chemistry
Volume	78
Issue number	19
DOIs	https://doi.org/10.1021/ac060693n
State	Published - Oct 1 2006
Externally published	Yes

ASJC Scopus subject areas

Analytical Chemistry

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Emmett, M. R., Kazazic, S., Marshall, A. G., Chen, W., Shi, S. D. H., Bolaños, B., & Greig, M. J. (2006). Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis. Analytical Chemistry, 78(19), 7058-7060. https://doi.org/10.1021/ac060693n

Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis. / Emmett, Mark R.; 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 journal › Article

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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.",

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