Nano-LC FTICR tandem mass spectrometry for top-down proteomics: routine baseline unit mass resolution of whole cell lysate proteins up to 72 kDa.

Analytical Chemistry
Jeremiah D TiptonAlan G Marshall

Abstract

Current high-throughput top-down proteomic platforms provide routine identification of proteins less than 25 kDa with 4-D separations. This short communication reports the application of technological developments over the past few years that improve protein identification and characterization for masses greater than 25 kDa. Advances in separation science have allowed increased numbers of proteins to be identified, especially by nanoliquid chromatography (nLC) prior to mass spectrometry (MS) analysis. Further, a goal of high-throughput top-down proteomics is to extend the mass range for routine nLC MS analysis up to 80 kDa because gene sequence analysis predicts that ~70% of the human proteome is transcribed to be less than 80 kDa. Normally, large proteins greater than 50 kDa are identified and characterized by top-down proteomics through fraction collection and direct infusion at relatively low throughput. Further, other MS-based techniques provide top-down protein characterization, however at low resolution for intact mass measurement. Here, we present analysis of standard (up to 78 kDa) and whole cell lysate proteins by Fourier transform ion cyclotron resonance mass spectrometry (nLC electrospray ionization (ESI) FTICR MS). ...Continue Reading

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Citations

Sep 6, 2012·Journal of the American Society for Mass Spectrometry·Behrooz Zekavat, Touradj Solouki
Feb 6, 2014·Journal of the American Society for Mass Spectrometry·Scott B FicarroJarrod A Marto
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Aug 10, 2012·Analytical Chemistry·Jorge Fernandez-de-Cossio Diaz, Jorge Fernandez-de-Cossio
Feb 27, 2015·Analytical Chemistry·Simone NicolardiYuri E M van der Burgt

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