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  4. Mesoporous TiO2-Based Experimental Layout for On-Target Enrichment and Separation of Multi- and Monophosphorylated Peptides Prior to Analysis with Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
 
research article

Mesoporous TiO2-Based Experimental Layout for On-Target Enrichment and Separation of Multi- and Monophosphorylated Peptides Prior to Analysis with Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry

Eriksson, Anna
•
Bergquist, Jonas
•
Edwards, Katarina
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2011
Analytical Chemistry

A simple method for on-target enrichment and subsequent sepn. and anal. of phosphorylated peptides is presented. The tryptic digest of a phosphorylated protein, in this case β-casein, is loaded onto a spot on a thin stripe made of mesoporous TiO2 sintered onto a conductive glass surface. After washing with a salicylic buffer to remove the nonphosphorylated peptides, the stripe is placed in an elution chamber contg. a phosphate soln. In a way analogous to thin layer chromatog. (TLC), the phosphate soln. acts as an eluent, clearly sepg. multi- and monophosphorylated peptides. By performing matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS) along the stripe, the detection of all phosphorylated peptides present in the digest is facilitated, as they are isolated from each other. The method was also tested on com. drinking milk, achieving successful sepn. between multi- and monophosphorylated peptides, as well as a detection limit in the femtomole range. As the enrichment, sepn., and anal. take place in the same substrate, sample handling and risk of contamination and sample loss is minimized. The results obtained suggest that the method, once optimized, may successfully provide a complete phosphoproteome.

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Type
research article
DOI
10.1021/ac1027879
Author(s)
Eriksson, Anna
Bergquist, Jonas
Edwards, Katarina
Hagfeldt, Anders  
Malmstrom, David
Agmo Hernandez, Victor.
Date Issued

2011

Published in
Analytical Chemistry
Volume

83

Start page

761

End page

766

Subjects

mesoporous titanium dioxide target phosphopeptide enrichment MALDI mass spectrometry

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
LSPM  
Available on Infoscience
July 6, 2015
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/115814
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