Pioneering continuous cell disruption for efficient commercial scale virus-like particle (VLP) biomanufacturing

Project Summary/Abstract Vaccines for many life threatening diseases have not been available to numerous global beneficiaries due to high product costs and limited manufacturing output capacities by commercial suppliers. The commercial human papilloma virus (HPV) vaccine produced using yeast, similar to vaccines for other infectious diseases, is based on the use of virus-like particles (VLPs) owing to their profound antigenicity and documented safety. The current industry standard practices for VLP manufacturing requires many individual processing steps with substantial human oversight, largely driven by commercial manufacturers' need to extract VLPs from inside the host organisms. Today, there is not a simple, automated method to achieve robust yields of VLPs from biomanufacturing processes, which in turn leads to high costs of goods manufactured (COGsM) for this critical vaccine format. The need to simplify VLP vaccine manufacturing to lower COGsM presents an opportunity to implement a first-in-class automated, continuous cell disruption method integrated with a continuous cell culturing bioreactor platform for VLP vaccine manufacturing. This Fast-Track SBIR proposes the development of a strategy, including a fully-automated equipment approach, for the cost effective recovery of VLPs from commercially used biomanufacturing hosts, like yeast. Our method relies on the efficient continuous disruption of cells to liberate the internally expressed VLPs produced during fermentation using our existing commercial bioreactor product, the Daisy Petal™ Perfusion Bioreactor System. We will use our expertise in building first-in-class yeast biomanufacturing hardware solutions and our inducible cell lysis platform to build a continuous disruption module for VLP bioproduction of an HPV VLP clinical candidate. Phase I will establish the commercial potential of continuous yeast cell disruption for VLP production. Phase I Aim 1 will demonstrate that self-assembled VLPs can be released and isolated from yeast cells via inducible cell disruption compatible with continuous operation. Phase II will build and demonstrate a hardware prototype for deployment of the novel continuous disruption method. Phase II Aim 1 will optimize engineered Pichia pastoris for inducible cell disruption and demonstrate utility for multiple VLP products. Phase II Aim 2 will build and demonstrate a GMP-ready prototype for continuous yeast cell disruption. After validating the commercial continuous disruption platform using the HPV VLP vaccine, VLP vaccine manufacturers can implement this new platform into their manufacturing process to increase manufacturing capacity, lower COGsM, and prevent manufacturing disruptions to improve the accessibility of efficacious vaccine products for patients worldwide. This anticipated commercial product will serve the urgent need to improve VLP biomanufacturing processes to address preventable diseases like HPV and HBV, and avert future VLP vaccine shortages.