(493c) Use of High-Throughput Purification to Support Definition of An Upstream Final Manufacturing Process for a Novel Vaccine

A High-throughput purification (downstream) method is required to support the fermentation (upstream) process development and pre-process validation activities for a novel vaccine, which must be completed within a compressed timeline. Purification of the upstream cell slurry is required to link quality attributes of the antigen to the fermentation operating parameters. In order to meet timelines, a high throughput purification method was developed capable of purifying 650 fermentation conditions in 8 months to support upstream process development and pre-process validation activities. This high throughput purification process utilizes mechanical lysis with silica beads and clarification by centrifugation. The clarified lysate is further purified by automated chromatography steps on a Tecan Freedom Evo® workstation using PhyTip® disposable pipette tips pre-filled with microliter volumes of resin. The high-throughput process offers two major advantages over lab-scale purification: less input material and time. The smallest lab scale purification requires a minimum of 110 g wet cell weight (wcw) input and demands two full time operators for 40 months to purify 650 fermentation conditions. In addition, the required input has potential impact on the fermentation if several timepoint samples require purification. The high-throughput purification process reduces input 100 fold and increases the throughput 10 fold. This savings also accounts for duplicate sample processing to increase the reliability of the purification data. In addition, the values of three quality attributes of the antigen purified by the high throughput method correlate within 15% of the values achieved at lab scale. As a result, 69,000 g wcw input and 32 months of employee time are saved, and all timelines are met by using this high-throughput purification method.