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 | |
| State | Published - Oct 1 2006 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis
AU - Emmett, Mark
AU - Kazazic, Sasa
AU - Marshall, Alan G.
AU - Chen, Wei
AU - Shi, Stone D H
AU - Bolaños, Ben
AU - Greig, Michael J.
PY - 2006/10/1
Y1 - 2006/10/1
N2 - 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.
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.
UR - http://www.scopus.com/inward/record.url?scp=33749490463&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33749490463&partnerID=8YFLogxK
U2 - 10.1021/ac060693n
DO - 10.1021/ac060693n
M3 - Article
VL - 78
SP - 7058
EP - 7060
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 19
ER -