Format

Send to

Choose Destination
Clin Chim Acta. 2014 Jul 1;434:16-20. doi: 10.1016/j.cca.2014.04.003. Epub 2014 Apr 18.

Comparative evaluation of peptide desalting methods for salivary proteome analysis.

Author information

1
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: nico.jehmlich@ufz.de.
2
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: claas@golatowski.de.
3
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: annette.murr@uni-greifswald.de.
4
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: gesell@uni-greifswald.de.
5
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: dhoplevm@uni-greifswald.de.
6
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: hammer@uni-greifswald.de.
7
Department of Functional Genomics, Interfaculty Institute for Genetics and Functional Genomics, University Medicine Greifswald, Germany. Electronic address: voelker@uni-greifswald.de.

Abstract

BACKGROUND:

Reliability and reproducibility are common requirements for high-quality generation of proteome data using mass spectrometry. The aim of this study was to compare four single-step desalting devices to provide a reproducible, high-recovery method for concentrating and purifying tryptic peptides before LC-MS/MS measurements.

MATERIAL AND METHODS:

Four different methods for peptide purification prior LC-MS/MS analyses (μC18 ZipTip® pipette tips, C18 ZipTip® pipette tips, TopTip C-18 and OASIS® HLB μElution Plate) were tested using whole saliva from healthy volunteers. A number of protein identifications and salivary protein patterns were analyzed comparatively.

RESULTS:

Each desalting device facilitated the identification of about 340 proteins. Purification-method dependent variations in protein composition were observed. Nevertheless, the overall inter-approach Pearson correlation coefficients of >0.95 indicate high reproducibility, reliability and recovery of proteins.

CONCLUSION:

The applied devices performed equally well in the removal of low molecular weight contaminants and provide high-quality data for quantitative proteomic analysis. Thus, selection should be primarily based on the amount of peptide extract available and the number of samples to be processed.

KEYWORDS:

Peptide desalting devices; Peptide purification; Saliva proteomics; Shotgun LC–MS/MS

PMID:
24751662
DOI:
10.1016/j.cca.2014.04.003
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center