A Powerful Purification Platform Technology We Did Not Talk About: Hydrophobic Interaction Chromatography
Hydrophobic interaction chromatography (HIC) separates molecules based on differences in their hydrophobicity. It is an important technique in both sample analysis and preparative scale purification. This post focuses on general information about HIC applications in bioprocessing. Across the industry, it is mainly used to remove both product-related impurities, such as aggregates, and process-related contaminants, such as host cell proteins.¹
HIC columns have been used in the purification of antibodies, recombinant proteins, and plasmid DNA. Lately, there are also literatures reporting using HIC columns for mRNA purification, with HIC columns being specifically well-suited for the polishing step.²
Ligands on HIC columns usually contain functional groups; the most common ones include alkyl and aryl groups, such as butyl (four carbons), octyl (eight carbons), and phenyl groups.³
Under certain conductivities and salt combinations, the hydrophobic region of the target molecule will be exposed and interact with the hydrophobic moieties. Depending on the specific types of separation targets and HIC columns, the loading buffer composition can be optimized to achieve better purification. Low conductivity buffers are usually used to elute the target from HIC columns.
Membrane chromatography columns can operate at much higher flow rates than resin chromatography columns. Typically, the membrane has lower binding capacity than resins. However, Purilogics offers HIC membranes with much higher binding capacity than leading HIC resin products, particularly for nucleic acids, such as plasmid DNA and RNA.
We also offer HIC membrane with different moieties with varying levels of hydrophobicity, from low to high.
If you are interested in our HIC membranes, please contact us at info@purilogics.com or +1 (864) 455-1457.
References
1. Lu, Y.; Williamson, B.; Gillespie, R. Recent Advancement in Application of Hydrophobic Interaction Chromatography for Aggregate Removal in Industrial Purification Process. Curr Pharm Biotechnol 2009, 10 (4), 427–433.2. Gagnon, P.; Goricar, B.; Persic, S.; Cernigoj, U.; Strancar, A. Two new capture options for improved purification of large mRNA. BioInsights; 2020.3. Mahn, A. Hydrophobic Interaction Chromatography: Fundamentals and Applications in Biomedical Engineering; 2012.