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Title: Evaluation of SDS depletion using an affinity spin column and IMS-MS detection

Abstract

While the use of detergents is necessary for a variety of protein isolation preparation protocols, often prior to mass spectral (MS) analysis, they are not compatible with MS analysis due to ion suppression and adduct formation. This manuscript describes optimization of detergent removal, using commercially available SDS depletion spin columns containing an affinity resin, providing for both increased protein recovery and thorough SDS removal. Ion mobility spectrometry coupled with mass spectrometry (IMS-MS) allowed for a concurrent analysis of both analyte and detergent. In the case of both proteins and peptides, higher detergent concentrations than previously reported provided an increase of sample recovery; however there was a limit as SDS was detected by IMS-MS at higher levels of SDS indicating incomplete detergent depletion. The results also suggest optimal conditions for SDS removal are dependent on the sample concentration. Overall, this study provides a useful guide for proteomic studies where SDS is required for efficient sample preparation.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
1060124
Report Number(s):
PNNL-SA-87924
40072; KP1704020
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Proteomics, 12(21):3138-3142
Additional Journal Information:
Journal Name: Proteomics, 12(21):3138-3142
Country of Publication:
United States
Language:
English
Subject:
Environmental Molecular Sciences Laboratory

Citation Formats

Hengel, Shawna M., Floyd, Erica A., Baker, Erin Shammel, Zhao, Rui, Wu, Si, and Pasa-Tolic, Ljiljana. Evaluation of SDS depletion using an affinity spin column and IMS-MS detection. United States: N. p., 2012. Web. doi:10.1002/pmic.201200168.
Hengel, Shawna M., Floyd, Erica A., Baker, Erin Shammel, Zhao, Rui, Wu, Si, & Pasa-Tolic, Ljiljana. Evaluation of SDS depletion using an affinity spin column and IMS-MS detection. United States. doi:10.1002/pmic.201200168.
Hengel, Shawna M., Floyd, Erica A., Baker, Erin Shammel, Zhao, Rui, Wu, Si, and Pasa-Tolic, Ljiljana. Thu . "Evaluation of SDS depletion using an affinity spin column and IMS-MS detection". United States. doi:10.1002/pmic.201200168.
@article{osti_1060124,
title = {Evaluation of SDS depletion using an affinity spin column and IMS-MS detection},
author = {Hengel, Shawna M. and Floyd, Erica A. and Baker, Erin Shammel and Zhao, Rui and Wu, Si and Pasa-Tolic, Ljiljana},
abstractNote = {While the use of detergents is necessary for a variety of protein isolation preparation protocols, often prior to mass spectral (MS) analysis, they are not compatible with MS analysis due to ion suppression and adduct formation. This manuscript describes optimization of detergent removal, using commercially available SDS depletion spin columns containing an affinity resin, providing for both increased protein recovery and thorough SDS removal. Ion mobility spectrometry coupled with mass spectrometry (IMS-MS) allowed for a concurrent analysis of both analyte and detergent. In the case of both proteins and peptides, higher detergent concentrations than previously reported provided an increase of sample recovery; however there was a limit as SDS was detected by IMS-MS at higher levels of SDS indicating incomplete detergent depletion. The results also suggest optimal conditions for SDS removal are dependent on the sample concentration. Overall, this study provides a useful guide for proteomic studies where SDS is required for efficient sample preparation.},
doi = {10.1002/pmic.201200168},
journal = {Proteomics, 12(21):3138-3142},
number = ,
volume = ,
place = {United States},
year = {2012},
month = {11}
}