Hydrophobic Interaction Chromatography for Bottom-Up Proteomics Analysis of Single Proteins and Protein Complexes

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 separ...

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Bibliographic Details
Published in:Journal of proteome research 2017-06, Vol.16 (6), p.2318-2323
Main Authors: Rackiewicz, Michal, Große-Hovest, Ludger, Alpert, Andrew J, Zarei, Mostafa, Dengjel, Jörn
Format: Article
Language:English
Subjects:
Cell Line
Chromatography, Liquid
Chromatography, Reverse-Phase - methods
Humans
Hydrophobic and Hydrophilic Interactions
Multiprotein Complexes - analysis
Proteins - analysis
Proteomics - methods
Tandem Mass Spectrometry
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Summary: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.
ISSN:1535-3893
1535-3907
DOI:10.1021/acs.jproteome.7b00015