Hydrophobic Interaction Chromatography (HIC)
Today’s bioprocessing requires more precision and more efficient operations. Hydrophobic interaction chromatography (HIC) separates molecules based on differences in their surface hydrophobicity. This technology is widely used in chromatographic purification, especially as a more precise purification step from other methods like ion exchange (IEX) or affinity. HIC typically requires high salt concentration for efficient binding of the target protein. Therefore, it is ideally used after chromatographic steps where elution is accomplished with high salt concentrations, i.e. ion exchange chromatography.
Sartorius has a range of HIC modalities (resins, membranes, and monoliths) to match your application.
Powerful method for removing aggregates
Ideal after initial sample cleanup and IEX chromatography
Ideal for final polishing steps or intermediate large-molecule purification
Find the Right Solution for Your Hydrophobic Interaction Chromatography Process
HIC is primarily used for aggregate removal due to its superior performance in that function.
Resins exhibit good binding capacities for small and medium-size proteins, while mixed-mode resins often combine HIC and IEX mechanisms.
Membranes have a higher binding capacity to viruses, virus-like particles, and other large complexes compared to resins, and also offer the advantage of higher flow rates resulting in shorter process time and higher productivity.
HIC monoliths are key in separating plasmid DNA isoforms.
Application | Capture of large biomolecules, polishing of all biomolecules | Capture and polishing of large biomolecules |
Implementation | Intrabatch multi-use, single-use between batches | Intrabatch multi-use, single-use between batches |
Ease of use | Ready-to-use | Ready-to-use |
Scalability | Capsules and cassettes for linear scale-up | Wide range of device sizes |
Sartobind® Membranes in HIC
Sartobind® Phenyl – Easy Aggregate Removal
Sartobind® Phenyl is a hydrophobic interaction membrane with low ligand substitution. This allows for mild elution conditions for the purification of all biomolecules.
Sartobind® Phenyl membranes can be considered as a replacement to columns for polishing (flow-through) operations and a number of bind-and-elute applications, as they work at much higher flow rates, reduced complexity and without size exclusion effects when purifying large biomolecules.
The typical dynamic binding capacity (DBC) at 10 % breakthrough is:
• 9-13 mg/mL mAb
• 10 mg/mL BSA
• 23 mg/mL Lysozyme
Sartobind® Phenyl Mini
The new Sartobind® Phenyl Mini provides 20 mL membrane volume, which allows bioprocess customers easier scale-up and is a perfect fit for the production of diagnostic products. Sartobind® Mini is directly scalable with the existing Sartobind portfolio which ranges from process development (Nano) to commercial manufacturing (Jumbo and cassettes).
Monoliths for HIC
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 adenoassociated viruses (AAV).
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) |
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CIMmultus® OH – Hydrophobic Interaction Chromatography
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 - Hydrophobic Interaction Chromatography
Strongly hydrophobic, (uncharged) ligand
Working range, pH 2–10
Highly effective for removing proteins from nucleic acids (pDNA, RNA)
