Hydrophobic Interaction Chromatography

Thermo Fisher Scientific provides a suite of POROS HIC resins offering a differentiating range in hydrophobicity suitable for bind/elute and flow-through applications. The resins can be utilized at lower salt concentrations for the purification of a wide variety of biomolecules including therapeutic proteins, antibody fragments, antibody drug conjugates (ADCs), and other large molecules.  

	POROS HIC Resin	Surface Chemistry	Key Application
	POROS Ethyl	Novel ethyl	Bind/elute mode of moderately to considerably hydrophobic molecules
	POROS Benzyl	Low density benzyl/aromatic	Bind/elute or flow-through mode depending on molecule
	POROS Benzyl Ultra	High density benzyl/aromatic	Flow-through mode in lower salt to bind impurities such as aggregates

Figure 1: Order of hydrophobicity of POROS HIC resins. Hydrophobicity based on lysozyme gradient elution. 

The new 50 µm resin backbone consists of cross‐linked poly(styrene-divinylbenzene) with a unique pore structure that provides rapid mass transport and enables enhanced productivity. The particle surface is coated with a novel polymer coating, which is then further derivatized with a range of hydrophobic ligands for flexible purification process design (Figure 2). POROS HIC resins deliver superior performance independent of flow rate, a major improvement in the purification tool box for impurity removal.

• Differentiating selectivity and superior resolution capability which helps improve yield and purity, reduce cost of goods (COG)
• High capacity for a range of molecules leading to reduced column size and smaller footprint
• Use of lower salt concentrations and weaker lyotropic salts allowing for increased process optimization flexibility with lower conductivity while maintaining superior resolution
• Flow rate independent performance
• Robust stability

Figure 2. Separation of a mixture of standard proteins using POROS Ethyl, POROS Benzyl and POROS Benzyl Ultra resins. Column size: 0.46cmD X 20cmL; Protein Mixture: Ribonuclease A, Lysozyme, Chymotrypsin and Chymotrypsinogen in 1.7M ammonium sulfate, 50mM sodium phosphate pH 7.0; Gradient Elution: 1.7M ammonium sulfate/50 mM sodium phosphate pH 7.0 to 50 mM sodium phosphate pH 7.0 over 10 column volumes (CV); Flow rate: 100 cm/hr.

For research use only. Not for use in diagnostic procedures.