Low-molecular-weight color pI markers to monitor on-line the peptide focusing process in OFFGEL fractionation

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081715%3A_____%2F17%3A00477435" target="_blank" >RIV/68081715:_____/17:00477435 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1002/elps.201700075" target="_blank" >http://dx.doi.org/10.1002/elps.201700075</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/elps.201700075" target="_blank" >10.1002/elps.201700075</a>

Result language

angličtina

Original language name

Low-molecular-weight color pI markers to monitor on-line the peptide focusing process in OFFGEL fractionation

Original language description

High-throughput mass spectrometry-based proteomic analysis requires peptide fractionationnto simplify complex biological samples and increase proteome coverage. OFFGELnfractionation technology became a common method to separate peptides or proteins usingnisoelectric focusing in an immobilized pH gradient. However, the OFFGEL focusingnprocess may be further optimized and controlled in terms of separation time and pInresolution. Here we evaluated OFFGEL technology to separate peptides from differentnsamples in the presence of low-molecular-weight (LMW) color pI markers to visualizenthe focusing process. LMW color pI markers covering a large pH range were added tonthe peptide mixture before OFFGEL fractionation using a 24-wells device encompassingnthe pH range 3–10. We also explored the impact of LMW color pI markers on peptidenfractionation labeled previously for iTRAQ. Then, fractionated peptides were separated bynRP_HPLC prior to MS analysis using MALDI-TOF/TOF mass spectrometry in MS andnMS/MS modes. Here we report the performance of the peptide focusing process in thenpresence of LMW color pI markers as on-line trackers during the OFFGEL process andnthe possibility to use them as pI controls for peptide focusing. This method improves thenworkflow for peptide fractionation in a bottom-up proteomic approach with or withoutniTRAQ labeling.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10406 - Analytical chemistry

Project

—

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Electrophoresis

ISSN

0173-0835

e-ISSN

—

Volume of the periodical

38

Issue of the periodical within the volume

16

Country of publishing house

DE - GERMANY

Number of pages

8

Pages from-to

2034-2041

UT code for WoS article

000408038500008

EID of the result in the Scopus database

2-s2.0-85026328714