What Are the Applications of Hydrophilic Interaction Liquid Chromatography in Proteomics
Hydrophilic Interaction Liquid Chromatography (HILIC), a variant of High-Performance Liquid Chromatography (HPLC), is primarily employed for the separation of polar compounds. Owing to its unique separation mechanism, HILIC has emerged as a powerful tool in proteomic analysis and the investigation of complex biological samples.
In the context of proteomics, HILIC finds application in the following key areas:
1. Peptide Enrichment
HILIC enables efficient enrichment and separation of polar peptides within biological samples. This is particularly beneficial for subsequent mass spectrometry (MS) analysis, as it reduces sample complexity and enhances the detectability of peptides present at low abundance.
2. Glycoprotein Separation
Due to the high polarity of glycosylated peptides, HILIC is well-suited for glycoproteomic studies, assisting researchers in characterizing the extent and types of protein glycosylation.
3. Coupling with Mass Spectrometry
HILIC is often integrated with MS techniques, offering enhanced sensitivity and resolution—especially valuable in the analysis of proteins bearing multiple post-translational modifications and complex peptide sequences.
4. Pre-Fractionation Step
Within multidimensional liquid chromatography (MDLC) strategies in proteomics, HILIC serves as an effective pre-fractionation approach, facilitating improved separation efficiency in downstream analyses by reducing sample complexity.
Despite its advantages, the application of HILIC in proteomics faces certain challenges, including limited analyte recovery, issues with reproducibility, and complexities in method development. These limitations necessitate optimization of operating conditions and the implementation of standardized protocols.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
Related Services
How to order?
