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

作者：Rackiewicz Michal; Grosse Hovest Ludger; Alpert Andrew J; Zarei Mostafa^*; Dengjel Joern^*

来源：Journal of Proteome Research, 2017, 16(6): 2318-2323.

DOI：10.1021/acs.jproteome.7b00015

• antibody
• protein complex
• chromatography
• mass spectrometry
• peptide
• bottom-up proteomics
• enzyme
• biologic

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.

• 出版日期2017-6