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

Kód výsledku v 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>

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2017

Kód důvěrnosti údajů

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

Název periodika

Electrophoresis

ISSN

0173-0835

e-ISSN

—

Svazek periodika

38

Číslo periodika v rámci svazku

16

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

8

Strana od-do

2034-2041

Kód UT WoS článku

000408038500008

EID výsledku v databázi Scopus

2-s2.0-85026328714