Although hydrophobic interaction chromatography (HIC) is an established production method for large-scale protein purification, the need to add lyotropic salts during binding and elution makes it both costly and environmentally challenging.
However, researchers from the >Pall Corporation, have presenting data on a new mixed-mode chromatography method combining hydrophobic and ionic components that give process chromatographers versatility in protein purification,
The new Pall BioSepra PPA HyperCel and HEA HyperCel mixed-mode chromatography sorbents are capable of interacting with a range of proteins under physiological-type conditions (pH and ionic strength) without the need to add lyotropic salts at concentrations typical of those required for traditional HIC.
The added lyotropes require disposal following the adsorption and wash stages, resulting in additional expense beyond initial reagent costs, and can also significantly impact the environment, especially with large-scale production.
"HEA HyperCel and PPA HyperCel add universal protein purification tools to the Pall process chromatography toolbox," said Peter Levison, Technology Development director.
"These sorbents answer the need for innovation in downstream processing, the source of most of today's biopharmaceutical manufacturing bottlenecks due to increasingly larger yields produced upstream."
The combined characteristics of HIC and ion exchange chromatography (IEX) in the new sorbents allow process chromatographers to adjust modalities to capture virtually any protein.
The data demonstrated the binding capacity of the new sorbents compared to conventional HIC sorbents.
The study evaluated the binding and elution properties of PPA and HEA HyperCel with a variety of proteins and more complex feedstreams.
It also included a comparison of PPA and HEA HyperCel with anion exchange, HIC and HCIC sorbents.
The effect of temperature on adsorption of BSA and ovalbumin was also investigated. It was found that protein-binding capacity using the new sorbents increases as temperatures rise, demonstrating a similarity to HIC processes, even in the absence of lyotropes.
"Eliminating the need to add lyotropic salts provides a considerable economic and environmental benefit for chromatography operations," commented Levison.
"These sorbents also provide a higher yield and recovery under selected conditions."
The results of this study were presented as part of the scientific program at the 19th International PREP Symposia in Baltimore, Maryland.