The adoption of High Throughput (HT) chromatography techniques during downstream process development activities for large vaccine molecules is frequently hampered by the poor compatibility of conventional chromatography media with the large size of such biomolecules. Further challenges in implementing a HT vaccines process development workflow accrue by both the often large variability in typical virus titer assays, such as plaque assays, and their low throughput. With the advent of novel adsorbents, specifically designed to accommodate such molecules, it is important to increase the understanding on the performance of HT chromatography formats within the context of vaccines process development. Here, the filter plate and miniature column HT chromatography methods were employed to carry out scouting studies for a large live virus vaccine candidate. The results from these studies were compared against those obtained from conventional bench scale runs to establish the ability of these HT techniques to guide early phase process development activities. Binding studies were performed across all formats and scales, for membrane and porous media of ion exchange and multimodal functionalities. In these studies, the involved analytics included an automated 384 well plate TCID50 assay for product tracking, allowing for increased levels of replication while accommodating the large number of generated samples, and routinely deployed HT analytics for the determination of contaminant levels such as host cell DNA. Experiments were performed on automated liquid handling platforms enabling walk-away automation. The obtained results were analyzed with statistical tools to assess, quantitatively, the agreement between the emerging windows of operation and developed models across scales. It was shown that both, the filter plate and miniature column HT methods, delivered trends similar to those observed for their bench scale counterparts, albeit with a fraction of the experimental expenditures. Hence, both HT chromatography formats were capable of yielding valuable information for early phase vaccines process development activities. These results serve to cement further the value of High Throughput techniques in bioprocessing activities.

Assessment of automated high throughput chromatography techniques in vaccines process development

2019 Spring National Meeting

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