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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Hydrophilic interaction liquid chromatography (HILIC) in proteomics
Paul J Boersema1, Shabaz Mohammed, Albert J R Heck
1Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht, The Netherlands.
Analytical and Bioanalytical Chemistry
|February 12, 2008
Summary
Hydrophilic interaction liquid chromatography (HILIC) shows promise as an alternative to strong cation exchange (SCX) chromatography in multidimensional proteomics. HILIC offers enhanced separation power and improved analysis of post-translational modifications.
Area of Science:
- Proteomics
- Analytical Chemistry
- Chromatography
Background:
- Nanoflow multidimensional chromatography is the standard for peptide separation in proteomics.
- Strong cation exchange (SCX) and reversed-phase (RP) chromatography are commonly combined for peptide separation.
- Orthogonal separation characteristics are ideal for multidimensional chromatography.
Purpose of the Study:
- To review the potential of hydrophilic interaction liquid chromatography (HILIC) in multidimensional peptide separation for proteomics.
- To evaluate HILIC as an alternative to SCX chromatography.
- To highlight HILIC's advantages in separation power and post-translational modification analysis.
Main Methods:
- Review of recent scientific literature on HILIC in proteomics.
- Comparison of HILIC with SCX and RP chromatography for peptide separation.
- Analysis of HILIC's performance in multidimensional chromatography setups.
Main Results:
- HILIC demonstrates potential as a viable alternative to SCX chromatography.
- HILIC offers advantages in terms of separation power for complex peptide mixtures.
- HILIC facilitates targeted analysis of protein post-translational modifications.
Conclusions:
- HILIC is a promising technique for multidimensional peptide separation in proteomics.
- HILIC may surpass SCX in certain separation applications.
- Further research into HILIC-based multidimensional chromatography is warranted for advancing proteomics.
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