Protein A (ProA) chromatography is an affinity-based method that offers high selectivity and robust platform capability. In recent years, the use of alkali-stabilized ProA resin has enabled direct capture of mAb from dirtier feeds as a direct result of stronger cleaning agents that can be effectively used to remove contaminants from a fouled ProA resin. However, these caustic and alkaline conditions need to be optimized in order to minimize loss of ProA ligand. Furthermore, there are no universal conditions that may be implemented, as cell culture and harvest conditions differ at each company. Traditional approaches to resin lifetime studies put a significant burden on both time and material resources. High-throughput and miniaturization of these studies can therefore be a powerful tool to explore wider ranges of conditions. To this end, a fully automated screening method was developed on the TECAN liquid handling system to study resin lifetime. This abstract outlines the process and results obtained for a mAb using MabSelect Sure<sup>Ò</sup> resin and 96-well filter plates. The biggest challenge to using filter plates is gaining consistent results for a sufficient number of cycles. In this study 16 different conditions on one plate were compared to determine cleaning conditions based on low product carry-over and high yields, with a method that delivered reliable results up to 25 cycles. To further screen the cleaning conditions, both fresh and stressed (fouled) resin were utilized to determine which condition best balanced high cleaning-ability and minimum ligand loss, thus maximizing the total number of allowable cleaning cycles. The established HT method can therefore be used to quickly optimize cleaning conditions for any mAb, and the selected conditions can then be verified by running scale-down model columns.

Development of a fully automated high-throughput (HT) method to screen cleaning conditions during a resin lifetime study

255th National Meeting (2018)

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