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 › peer-review

33 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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title = "Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis",

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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AU - Emmett, Mark R.

AU - Kazazic, Sasa

AU - Marshall, Alan G.

AU - Chen, Wei

AU - Shi, Stone D.H.

AU - Bolaños, Ben

AU - Greig, Michael J.

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

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