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
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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
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- English
- Classification
- Biological sciences
- License
- License undefined
- Identifiers
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- RERO DOC 288892
- DOI 10.1021/acs.jproteome.7b00015
- Persistent URL
- https://folia.unifr.ch/unifr/documents/305515
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