Protein Aggregates and Gel Filtration Chromatography: Improving Quantitation and Throughput of a UHPLC Method
Gel filtration chromatography (GFC) is the most widely used method for quantitating protein aggregates in therapeutic drugs. It is a simple method, but prone to error as a result of poor method development and column selection. GFC columns tend to non-specifically adsorb large proteins and aggregates resulting in poor quantitation of “true” aggregate amount. Sample “priming” and mobile phase optimization can help reduce such irregularities. Simple method development rules using new column technologies are presented that demonstrate improved accuracy for these methods.