Reduction and control of soluble protein aggregates present persistent challenges during process development of monoclonal antibody-based therapeutics, and a myriad of chromatographic techniques are available to aid in this effort. As with many of its peers, Takeda Pharmaceuticals utilizes a platform purification approach for early-stage programs, employing Protein A capture chromatography, cation exchange (CEX) chromatography, and anion exchange chromatography, where the bind-elute CEX step removes soluble aggregates, among other impurities. While such platform processes are robust and scalable, multiple bind-elute steps result in long process times, significant buffer consumption, and potential process bottlenecks. As cell line productivity continues climbing and the industry demands faster, cheaper purification operations, we encounter a dilemma: How do process scientists adapt platform operations to process large quantities of antibody, while meeting strict purity standards, especially for highly impure feeds?<br/><br/>The present work is a case study in platform purification development at Takeda. During development of a new cell line for an early-stage monoclonal antibody, the soluble aggregate content of the resulting antibody after harvest was nearly three-fold greater than that in the original cell line process. Despite optimization, the platform CEX process produced eluate with aggregate levels nearly twice those of the original cell line process.<br/><br/>To address this challenge, Takeda identified a novel hydrophobic interaction chromatography (HIC) resin whose high hydrophobicity permits aggregate removal under low-salt conditions and while operating in flow-through mode. Using small, pre-packed columns and design of experiment techniques, the process parameters were optimized to achieve robust aggregate clearance and high recovery, even at column loadings &gt; 100 g/L. Moreover, while the HIC step alone demonstrates significant aggregate removal capabilities, placement of an optimized CEX step immediately downstream shows further aggregate clearance. The optimized HIC step is therefore a powerful platform aggregate removal option, especially for challenging feeds with strict purity standards.

Optimization of a platform hydrophobic interaction chromatography step for robust aggregate removal in flow-through mode

2019 Spring National Meeting

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.