PXD001333

PXD001333 is an original dataset announced via ProteomeXchange.

Title	Anion-Exchange Chromatography of Tryptic and Phosphopeptides: WAX vs. SAX and AEX vs. ERLIC
Description	At a pH > 5, phosphopeptides have two negative charges per residue and are well-retained in anion-exchange chromatography. However, the peptides with one or two phosphate groups are not separated from the peptides with multiple Asp or Glu residues, which interfere with the identification of phosphopeptides in tryptic digests. At a pH around 2, phosphate residues have just a single negative charge but Asp and Glu have none. This facilitates the separation of phosphopeptides from unmodified acidic peptides. The singly phosphorylated peptides are retained too weakly under these conditions, due to electrostatic repulsion, unless hydrophilic interaction is superimposed in the ERLIC mode (electrostatic repulsion-hydrophilic interaction chromatography). Weak anion-exchange (WAX) and strong anion-exchange (SAX) columns were compared, both with peptide standards and with a complex tryptic digest. The SAX column exhibited higher capacity at pH 6 than did the WAX column. However, only about 60% as many phosphopeptides were identified at pH 6 than via ERLIC at pH 2. In one run, 12,467 phosphopeptides were identified, including 4233 with more than one phosphate. We conclude that chromatography of phosphopeptides is best performed at low pH in the ERLIC mode. Under those conditions the performances of the SAX and WAX materials were comparable.
HostingRepository	PRIDE
AnnounceDate	2015-04-23
AnnouncementXML	Submission_2015-04-23_01:21:27.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Otto Hudecz
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2014-09-17 01:20:55	ID requested
⏵ 1	2015-04-23 01:21:29	announced

Alpert AJ, Hudecz O, Mechtler K, Anion-exchange chromatography of phosphopeptides: weak anion exchange versus strong anion exchange and anion-exchange chromatography versus electrostatic repulsion-hydrophilic interaction chromatography. Anal Chem, 87(9):4704-11(2015) [pubmed]

curator keyword: Biomedical

submitter keyword: chromatography ERLIC SAX WAX AEX

Andrew Alpert
contact affiliation	PolyLC Inc., 9151 Rumsey Road, ste. 175, Columbia, MD 21045 USA
contact email	aalpert@polylc.com
lab head
Otto Hudecz
contact affiliation	Protein Chemistry Facility
contact email	hudecz@imp.univie.ac.at
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD001333
     1. Label: PRIDE project
     2. Name: Anion-Exchange Chromatography of Tryptic and Phosphopeptides: WAX vs. SAX and AEX vs. ERLIC