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The Use of Titanium Dioxide for Selective Enrichment of Phosphorylated Peptides.

Thingholm, Tine LU and Larsen, Martin R (2016) In Methods in Molecular Biology 1355. p.135-146
Abstract
Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2 chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice... (More)
Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2 chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice in large-scale phosphoproteomics. Here we describe a batch mode protocol for phosphopeptide enrichment using TiO2 chromatographic material followed by desalting and concentration of the sample by reversed phase micro-columns prior to downstream MS and LC-MS/MS analysis. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Methods in Molecular Biology
volume
1355
pages
135 - 146
publisher
Springer
external identifiers
  • pmid:26584923
  • scopus:84947976150
ISSN
1940-6029
DOI
10.1007/978-1-4939-3049-4_9
language
English
LU publication?
yes
id
aef55d3f-8c23-4150-bf88-6cded2ae3da0 (old id 8235109)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/26584923?dopt=Abstract
date added to LUP
2015-12-01 16:14:23
date last changed
2017-11-19 04:18:23
@article{aef55d3f-8c23-4150-bf88-6cded2ae3da0,
  abstract     = {Titanium dioxide (TiO2) has very high affinity for phosphopeptides and in recent years it has become one of the most popular methods for phosphopeptide enrichment from complex biological samples. Peptide loading onto TiO2 resin in a highly acidic environment in the presence of 2,5-dihydroxybenzoic acid (DHB), phthalic acid, lactic acid, or glycolic acid has been shown to improve selectivity significantly by reducing unspecific binding of non-phosphorylated peptides. The phosphopeptides bound to the TiO2 are subsequently eluted from the chromatographic material using an alkaline buffer. TiO2 chromatography is extremely tolerant towards most buffers used in biological experiments, highly robust and as such it has become the method of choice in large-scale phosphoproteomics. Here we describe a batch mode protocol for phosphopeptide enrichment using TiO2 chromatographic material followed by desalting and concentration of the sample by reversed phase micro-columns prior to downstream MS and LC-MS/MS analysis.},
  author       = {Thingholm, Tine and Larsen, Martin R},
  issn         = {1940-6029},
  language     = {eng},
  pages        = {135--146},
  publisher    = {Springer},
  series       = {Methods in Molecular Biology},
  title        = {The Use of Titanium Dioxide for Selective Enrichment of Phosphorylated Peptides.},
  url          = {http://dx.doi.org/10.1007/978-1-4939-3049-4_9},
  volume       = {1355},
  year         = {2016},
}