Hydrophobic Interaction Chromatography (HIC)
Modern bioprocessing demands greater precision and operational efficiency. Hydrophobic interaction chromatography (HIC) plays a key role in achieving this by separating molecules based on their surface hydrophobicity. Commonly used in downstream purification, HIC provides enhanced resolution and complements other techniques such as ion exchange (IEX) or affinity chromatography. Since HIC requires high salt concentrations to promote protein binding, it is particularly well-suited for use after steps like IEX, where proteins are typically eluted in high-salt buffers.
Sartorius offers a comprehensive portfolio of HIC solutions—including resins, membranes, and monoliths—designed to meet the specific needs of your application.
Hydrophobic Interaction Chromatography Portfolio
Find the Right Solution for Your HIC Process
HIC is a powerful tool primarily used for aggregate removal, thanks to its exceptional performance. Sartorius provides a versatile range of HIC solutions across resins, membranes, and monoliths to support diverse purification needs:
Membranes provide higher binding capacities for viruses, virus-like particles, and other large complexes, while also enabling faster flow rates for reduced processing time and improved productivity.
| Sartobind® Phenyl |
|---|---|
| Ligand | Phenyl (R-NH-C₆H₅) |
| Base Matrix | Cellulose |
| Focus Applications |
|
Resins offer strong binding capacities for small- and medium-size proteins, and mixed-mode resins combine HIC and IEX functionalities for enhanced selectivity.
| Methyl Ceramic Hyper D | SDR Hyper D® | |
|---|---|---|
| Ligand | Methyl-CH3 | C 11 |
| Base Matrix | Ceramic beads | Silica beads |
| Focus Applications |
|
|
The Choice for Hydrophobic Interaction Chromatography (HIC)
Hydrophobic interaction chromatography (HIC) is a must for nucleic acid separations. When combined with the advantages of monolithic chromatography, HIC meets this need while also providing an excellent solution for the purification of large biomolecules including adeno-associated viruses (AAV).
| CIMmultus® OH | CIMmultus® C4-HLD | |
|---|---|---|
| Channel sizes | 1.3, 2, 6 µm | 2, 6 µm |
| Column volume | 1 mL – 8 L | 1 mL – 8 L |
| Working range, pH | 2 – 10 | 2 – 10 |
| Functionality | Weak hydrophobic | Strong hydrophobic |
| Typical application | Viruses, (i.e. AAV), and VLPs | Nucleic acids (pDNA, mRNA) |
| Request a Quote | Request a Quote |
CIMmultus® OH
- Uncharged (neutral) ligand
- Working range, pH 2–10
- Retains very large solutes (virus particles, VLP, vesicles) in the presence of precipitating salts or polyethylene glycol
- Elutes large solutes with high resolution in order of increasing size
- Elution with descending salt or polyethylene glycol
- Sanitizable with 1 M NaOH
CIMmultus® C4-HLD
- Strongly hydrophobic (uncharged) ligand
- Working range, pH 2–10
- Highly effective for removing proteins from nucleic acids (pDNA, RNA)
Supporting Products & Services
Learn More About Hydrophobic Interaction Chromatography
Highlight Asset
Related Assets
Phenyl Membrane Adsorber for Bioprocessing - Sartobind® Hydrophobic Interaction Membrane Chromatography
PDF | 2.1 MB
Precision Purification for Diverse Bioprocessing Needs
Sartorius offers a broad and flexible portfolio of HIC tools designed to support the evolving requirements of modern bioprocessing. Whether the goal is to separate aggregates, complex viral particles, or plasmid isoforms, we offer HIC resins, membranes, and monoliths engineered for optimal performance. These solutions provide high binding capacities, fast flow rates, and advanced selectivity—including mixed-mode and salt-tolerant options—for scalable purification across a wide range of molecule sizes. The portfolio is designed to support application-specific process development with a focus on precision and adaptability.
Frequently Asked Questions
HIC is a technique for separating proteins and other biomolecules based on their surface hydrophobicity. This method is particularly useful for purifying proteins while maintaining their biological activity.
The four main process stages are the same compared to other chromatography techniques like ion exchange or mixed mode: equilibration, sample application and wash, elution, and regeneration.
Separation is done by applying a decreasing salt gradient in the mobile phase. The hydrophobic interactions between the mobile and stationary phases are weakened, allowing the molecules to elute from the column based on their hydrophobic properties.
Unlike traditional HIC columns, Sartobind® Phenyl membranes offer faster processing due to convective flow, lower buffer consumption, and easier scale-up thanks to single-use formats. They also do not require column packing.
Sartobind® Phenyl acts as a polishing tool to remove mAb aggregates and misfolded forms that are more hydrophobic, especially when ion exchange and Protein A alone are insufficient.
Binding typically requires moderate to high salt concentrations (e.g., 1–2 M ammonium sulfate), but Sartobind® Phenyl supports a broad salt tolerance and shows good binding characteristics across a wide conductivity range.
