A semi-virtual two dimensional gel electrophoresis: IF-ESI LC-MS/MS

Stanislav Naryzhny; Victor Zgoda; Artur Kopylov; Elena Petrenko; Аlexander Archakov

doi:10.1016/j.mex.2017.08.004

A semi-virtual two dimensional gel electrophoresis: IF-ESI LC-MS/MS

MethodsX. 2017 Aug 31:4:260-264. doi: 10.1016/j.mex.2017.08.004. eCollection 2017.

Authors

Stanislav Naryzhny^{1

2}, Victor Zgoda¹, Artur Kopylov¹, Elena Petrenko¹, Аlexander Archakov¹

Affiliations

¹ Orekhovich Institute of Biomedical Chemistry of Russian Academy of Medical Sciences, Pogodinskaya 10, Moscow, 119121, Russia.
² B.P. Konstantinov Petersburg Nuclear Physics Institute, National Research Center "Kurchatov Institute", Orlova roscha, Gatchina, Leningrad region, 188300, Russia.

Abstract

A method for increasing the productivity of ESI LC-MS/MS (electrospray ionization-liquid chromatography-tandem mass spectrometry) was proposed and applied. After IF (isoelectric focusing) of the sample using IPG (immobilized pH gradient) strip, the strip was cut to sections, and every section was treated according to trypsinolysis protocol for MS/MS analysis. The peptides produced were further analyzed by ESI LC-MS/MS. The procedure allows to: •identify many more proteins and proteoforms compared to shotgun analysis of extracts.•build a semi-virtual 2DE map of identified proteins.

Keywords: 2DE; A semi-virtual two dimensional gel electrophoresis: IF–ESI LC-MS/MS; IEF; Identification; Large-scale; Proteoforms; Shotgun.