Fully automated high-throughput process development for the novel purification of rotavirus vaccines
June 17-22, 2018
Downstream processing of biopharmaceuticals, such as immunoglobulins, recombinant proteins and protein-based vaccines, traditionally requires multiple purification steps that can introduce cost and time-related limitations. To avoid these, alternative purification strategies are sought involving novel and efficient processes. Mixed-mode chromatography based unit operations can achieve these by making use of their multimodal functionality. This adheres to the goal of the Ultra-low cost Transferable Automated Platform for Vaccine Manufacture (ULTRA) project and its ultimate aim to deliver vaccines with a cost of goods less than 15 cents per dose.
Herein, we show the application of mixed mode chromatography for the purification of a P2-VP8 (PATH) rotavirus antigen vaccine expressed in Pichia pastoris (MIT, MA, USA; J. Christopher Love). A high-throughput workflow was adopted for the development of the purification step which employed microscale chromatography with miniature columns. This was implemented on an automated liquid handling station (Tecan Evo 200). The performed scouting studies focused on combinations of different binding and elution conditions along with multiple resins and selectively accessing the ion exchange and hydrophobic integration mechanisms of the resins. Therefore, a wide range of conditions were assessed with walk-away automation. Conditions leading to high yields and purities were then scaled-up for verification purposes. This provided further evidence for the good scalability properties of the miniature chromatography column technique.
The results from this study demonstrate that mixed mode chromatography can potentially lead to the establishment of a highly desirable, one-step purification of rotavirus vaccine upon further optimisation. Hence, downstream processing in rotavirus vaccine production can be de-bottlenecked with systematic process development activities leading to significantly improved whole process efficiencies. The utility of this method is that it has generated reproducible and scalable data with reduced sample requirements to just a few millilitres very early on in the development process. More broadly, this high-throughput methodology will enable the early purification screening of multiple vaccine candidates and enable their selection for further development based on ease of processing.
Shaleem I. Jacob, Lourdes Velez, Tarit K. Mukhopadhyay, and Stephen A. Morris, "Fully automated high-throughput process development for the novel purification of rotavirus vaccines" in "Vaccine Technology VII", Amine Kamen, McGill University Tarit Mukhopadhyay, University College London Nathalie Garcon, Bioaster Charles Lutsch, Sanofi Pasteur Eds, ECI Symposium Series, (2018). http://dc.engconfintl.org/vt_vii/16