Issue 3, 2015

Direct analysis of in-gel proteins by carbon nanotubes-modified paper spray ambient mass spectrometry

Abstract

The in situ and direct extraction, desorption and ionization of in-gel intact proteins after electrophoresis has been achieved by carbon nanotubes (CNTs)-modified paper spray mass spectrometry at ambient conditions. Characteristics of CNTs (including larger surface area, smaller pore diameter and enhanced conductivity) were endowed to the porous filter paper substrate by uniformly dispersing the CNTs on the filter paper. Upon applying electric potential to the CNTs-modified paper, the in-gel proteins were extracted from the gel and subsequently migrated to the tip of the filter paper by electrophoresis-like behavior for paper spray ionization, which was monitored by extracted ion chronograms. The characterizations of modified filter papers and CNTs nanoparticles further confirmed the role of CNTs in in-gel protein extraction, protein migration as well as spray ionization at the paper tip. Under optimized conditions, a mixture of cytochrome c, lysozyme and myoglobin was successfully separated by native electrophoresis and subsequently analysed by the present method, showing a limit of detection of 10 ng per gel band. The present strategy offers a new pathway for the direct detection of in-gel intact proteins at ambient conditions without any pre-treatment (e.g. digestion, chemical extraction and desalting), which exhibits potential applications in top-down proteomics.

Graphical abstract: Direct analysis of in-gel proteins by carbon nanotubes-modified paper spray ambient mass spectrometry

Supplementary files

Article information

Article type
Paper
Submitted
15 Sep 2014
Accepted
07 Oct 2014
First published
08 Oct 2014

Analyst, 2015,140, 710-715

Author version available

Direct analysis of in-gel proteins by carbon nanotubes-modified paper spray ambient mass spectrometry

F. Han, Y. Yang, J. Ouyang and N. Na, Analyst, 2015, 140, 710 DOI: 10.1039/C4AN01688B

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