PXD006112

PXD006112 is an original dataset announced via ProteomeXchange.

Title	Improved reversed phase chromatography of hydrophilic peptides from spatial and temporal changes in column temperature
Description	Reversed phase chromatography is an established method for peptide separation and frequently coupled to electrospray ionization-mass spectrometry for proteomic analysis. Column temperature is one parameter that influences peptide retention and elution, but it is often overlooked as its implementation requires additional equipment and method optimization. An apparatus that allows temperature manipulation in three areas of a two-column setup was evaluated for improvements in chromatography. Using commercially available standards, we demonstrate that a low column temperature (0 °C) during sample loading enhances the peak shape of several bovine serum albumin hydrophilic peptides. For digested HeLa lysates, approximately 15% more peptide identifications were obtained by increasing the precolumn temperature to 50 °C after the 500 ng sample was loaded at a low temperature. This method also identified additional early eluting peptides with grand average of hydropathicity values less than -2. We also investigated the effect of cooler column temperatures on peptides with post-translational modifications. It was possible to minimize the co-elution of an isoaspartylated peptide and its unmodified version when the analytical column temperature was decreased to 5 °C. Aside from demonstrating the utility of lower temperatures for improved chromatography, its application at specific locations and time points is critical for peptide detection and separation.
HostingRepository	PRIDE
AnnounceDate	2017-04-10
AnnouncementXML	Submission_2017-04-10_00:23:10.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Clifford Young
SpeciesList	scientific name: Bos taurus (Bovine); NCBI TaxID: 9913; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; acetylated residue; modified L-aspartic acid residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-03-20 02:22:01	ID requested
⏵ 1	2017-04-10 00:23:11	announced

Young C, Podtelejnikov AV, Nielsen ML, Improved Reversed Phase Chromatography of Hydrophilic Peptides from Spatial and Temporal Changes in Column Temperature. J Proteome Res, 16(6):2307-2317(2017) [pubmed]

curator keyword: Technical

submitter keyword: BSA, HeLa, hydrophilic peptide, isoaspartic acid, liquid chromatography, mass spectrometry, temperature

Michael Lund Nielsen
contact affiliation	The Novo Nordisk Foundation Center for Protein Research, Proteomics Program, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200 Copenhagen, Denmark
contact email	michael.lund.nielsen@cpr.ku.dk
lab head
Clifford Young
contact affiliation	Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark.
contact email	clifford.young@cpr.ku.dk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD006112
     1. Label: PRIDE project
     2. Name: Improved reversed phase chromatography of hydrophilic peptides from spatial and temporal changes in column temperature