Separating Molecules by Hydrophobicity
Hydrophobic Interaction Chromatography (HIC) is a powerful technique for separating and purifying proteins based on their hydrophobicity. It’s particularly useful for sensitive biomolecules, as it employs relatively mild conditions compared to reversed-phase chromatography, allowing for the retention of protein tertiary structure and biological activity. In HIC, proteins are loaded onto a column with a hydrophobic stationary phase in the presence of high salt concentrations. As the salt concentration decreases, proteins elute in order of increasing hydrophobicity.
PolyLC’s HIC columns offer exceptional performance, enabling the separation and purification of proteins while preserving their native structures. This is crucial for applications like top-down proteomics and the analysis of proteins incompatible with reversed-phase conditions. Our columns are designed to minimize the salt concentration required for retention, making them compatible with direct analysis via mass spectrometry (MS).
In Hydrophobic Interaction Chromatography (HIC) proteins, in the presence of high salt concentration, will bind to the hydrophobic stationary phase then, as salt concentration decreases, proteins will elute in order of increasing hydrophobicity.
PolyLC HIC Columns: Optimized for Protein Separations
PolyLC offers a range of HIC columns with varying ligand sizes, allowing you to fine-tune selectivity and optimize separations for your specific needs. Unless a protein is known to be particularly hydrophobic, we recommend using PolyPROPYL A™.
Our HIC columns ensure:
- Preservation of Native Structure: Proteins maintain their tertiary structure and biological activity during HIC, making it ideal for sensitive biomolecules
- High Resolution: Achieve excellent separation of complex protein mixtures
- Compatibility with Various Sample Types: Analyze a wide range of proteins, including antibodies
FEATURED APPLICATION NOTE:
High-Resolution Analysis of Antibodies
PolyLC’s HIC columns excel in the analysis of antibodies, enabling the separation and characterization of glycoform variants with high resolution and sensitivity.
Further, the conditions achievable with PolyLC HIC columns allow for direct coupling with mass spectrometry, facilitating in-depth characterization of antibody structure and modifications.
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