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Title: Rapid Sample Processing For LC-MS Based Quantitative Proteomics Using High Intensity Focused Ultrasounds

Abstract

A new sample processing workflow that uses high intensity focused ultrasound to rapidly reduce and alkylate cysteines, digest proteins and then label peptides with 18O was developed for quantitative proteomics applications. Each step was individually refined to minimize reaction times, peptide loses and undesired by-products or modifications. By using this novel workflow, mouse plasma proteins were successfully denatured, alkylated, in-solution digested, and 18O labelled in < 10 min for subsequent analysis by liquid chromatography-electrospray ionization high resolution mass spectrometry. Performance was evaluated in terms of the number of mouse plasma peptides and proteins identified in a shotgun approach and the quantitative dynamic range. The results were compared with previously published results obtained using conventional sample preparation methods and were found to be similar. Advantages of the new method include greatly simplified and accelerated sample processing, as well as being readily amenable to automation.

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
947042
Report Number(s):
PNNL-SA-59400
KP1601010
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Proteome Research, 7(9):3860-3867; Journal Volume: 7; Journal Issue: 9
Country of Publication:
United States
Language:
English

Citation Formats

Lopez-Ferrer, Daniel, Heibeck, Tyler H., Petritis, Konstantinos, Hixson, Kim K., Qian, Weijun, Monroe, Matthew E., Mayampurath, Anoop M., Moore, Ronald J., Belov, Mikhail E., Camp, David G., and Smith, Richard D. Rapid Sample Processing For LC-MS Based Quantitative Proteomics Using High Intensity Focused Ultrasounds. United States: N. p., 2008. Web. doi:10.1021/pr800161x.
Lopez-Ferrer, Daniel, Heibeck, Tyler H., Petritis, Konstantinos, Hixson, Kim K., Qian, Weijun, Monroe, Matthew E., Mayampurath, Anoop M., Moore, Ronald J., Belov, Mikhail E., Camp, David G., & Smith, Richard D. Rapid Sample Processing For LC-MS Based Quantitative Proteomics Using High Intensity Focused Ultrasounds. United States. doi:10.1021/pr800161x.
Lopez-Ferrer, Daniel, Heibeck, Tyler H., Petritis, Konstantinos, Hixson, Kim K., Qian, Weijun, Monroe, Matthew E., Mayampurath, Anoop M., Moore, Ronald J., Belov, Mikhail E., Camp, David G., and Smith, Richard D. Mon . "Rapid Sample Processing For LC-MS Based Quantitative Proteomics Using High Intensity Focused Ultrasounds". United States. doi:10.1021/pr800161x.
@article{osti_947042,
title = {Rapid Sample Processing For LC-MS Based Quantitative Proteomics Using High Intensity Focused Ultrasounds},
author = {Lopez-Ferrer, Daniel and Heibeck, Tyler H. and Petritis, Konstantinos and Hixson, Kim K. and Qian, Weijun and Monroe, Matthew E. and Mayampurath, Anoop M. and Moore, Ronald J. and Belov, Mikhail E. and Camp, David G. and Smith, Richard D.},
abstractNote = {A new sample processing workflow that uses high intensity focused ultrasound to rapidly reduce and alkylate cysteines, digest proteins and then label peptides with 18O was developed for quantitative proteomics applications. Each step was individually refined to minimize reaction times, peptide loses and undesired by-products or modifications. By using this novel workflow, mouse plasma proteins were successfully denatured, alkylated, in-solution digested, and 18O labelled in < 10 min for subsequent analysis by liquid chromatography-electrospray ionization high resolution mass spectrometry. Performance was evaluated in terms of the number of mouse plasma peptides and proteins identified in a shotgun approach and the quantitative dynamic range. The results were compared with previously published results obtained using conventional sample preparation methods and were found to be similar. Advantages of the new method include greatly simplified and accelerated sample processing, as well as being readily amenable to automation.},
doi = {10.1021/pr800161x},
journal = {Journal of Proteome Research, 7(9):3860-3867},
number = 9,
volume = 7,
place = {United States},
year = {Mon Sep 01 00:00:00 EDT 2008},
month = {Mon Sep 01 00:00:00 EDT 2008}
}