410f High Throughput Protein Purification Using Negative Mode Hydrophobic Interaction Chromatography

Jennifer McNay, Kenneth Audo, and Kevin Bailey. Regeneron Pharmaceuticals, 777 Old Saw Mill River Road, Tarrytown, NY 10591

The biotechnology industry has seen a rapid increase in therapeutic protein production titers over the past few years. This has placed an increased demand on downstream purification operations to handle greater amounts of protein while maintaining high quality standards and process throughput. Flow-through or negative mode chromatography is commonly used in the bioprocess industry for anion exchange purification, and has the potential to dramatically increase process throughput. Work presented here examines the application of flow-through chromatography to aggregate removal for a large, heavily glycosylated protein using hydrophobic interaction chromatography (HIC). The effect of chromatographic operating parameters, including salt level, pH, and column loading, will be examined as a function of aggregate resolution. Scale-up considerations will also be examined to illustrate the potential benefit of this work to process throughput.