Hydrophobic interaction chromatography for bottom-up proteomics analysis of single proteins and protein complexes

Rackiewicz, Michal; Große-Hovest, Ludger; Alpert, Andrew J.; Zarei, Mostafa; Dengjel, Jörn

doi:10.1021/acs.jproteome.7b00015

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Hydrophobic interaction chromatography for bottom-up proteomics analysis of single proteins and protein complexes

Rackiewicz, Michal Department of Biology, University of Fribourg, Switzerland - Department of Dermatology, Medical Center - University of Freiburg, Germany
Große-Hovest, Ludger SYNIMMUNE GmbH, Tübingen, Germany
Alpert, Andrew J. PolyLC Inc., Columbia, United States
Zarei, Mostafa Solvias AG, Kaiseraugst, Switzerland
Dengjel, Jörn Department of Biology, University of Fribourg, Switzerland - Department of Dermatology, Medical Center - University of Freiburg, Germany

02.06.2017

Published in:

Journal of Proteome Research. - 2017, vol. 16, no. 6, p. 2318–2323

English Hydrophobic interaction chromatography (HIC) is a robust standard analytical method to purify proteins while preserving their biological activity. It is widely used to study post-translational modifications of proteins and drug–protein interactions. In the current manuscript we employed HIC to separate proteins, followed by bottom-up LC–MS/MS experiments. We used this approach to fractionate antibody species followed by comprehensive peptide mapping as well as to study protein complexes in human cells. HIC–reversed-phase chromatography (RPC)–mass spectrometry (MS) is a powerful alternative to fractionate proteins for bottom-up proteomics experiments making use of their distinct hydrophobic properties.

Faculty

Faculté des sciences et de médecine

Department

Département de Biologie

Language