Lu, Timothy Massachusetts Institute of Technology
To counter the current Ebola outbreak one treatment option involves a cocktail of three antibodies called 'ZMapp', which has been shown to cure primates and has been reportedly used in the current outbreak to treat a small number of people. Despite the promise of ZMapp, its use has been highly restricted to just a few patients because of its severely limited availability. This low availability is because ZMapp production is currently carried out in plants, a slow process that is difficult to expand rapidly. Other anti-Ebola antibodies are similarly constrained by production systems that are challenging, time-consuming, and expensive to engineer and scale-up. Thus, there is a tremendous need for generalizable platforms that can be rapidly engineered to produce anti-infectious drugs and then easily scaled-up to create large numbers of doses. In this RAPID proposal yeast strains optimized for rapid production of anti-infectious antibodies will be adapted to express anti-Ebola antibodies. Yeast are promising hosts for manufacturing therapeutic molecules because they can be transported without refrigeration, quickly grown to large scales, and modified to make humanized therapies. It is anticipate that the engineered yeasts will be useful for economical and large-scale manufacturing of anti-Ebola therapies. In addition, this work will provide rapid-response capabilities for tackling future emerging diseases because the yeast platform can be quickly engineered with synthetic biology tools to generate new therapeutic agents. In addition, a distributed biomanufacturing approach will be applied by coupling engineered yeast with a novel micro-bioreactor technology that has the potential to mount rapid responses at the source of disease outbreaks.
The goal of this RAPID proposal is to establish a rapid and flexible biomanufacturing platform in Pichia pastoris for the production of anti-Ebola neutralizing antibodies. ZMapp, a cocktail of 3 neutralizing monoclonal antibodies (mAbs), has been shown to rescue 100% of rhesus macaques when administered up to 5 days post-infection. Zmapp1 is currently produced in the plant Nicotiana benthamiana, a slow process that is difficult to scale. Current efforts to express neutralizing antibodies from other hosts, such as CHO cells, also require substantial time and expensive infrastructure to scale. Moreover, quick delivery and long-term storage of mAb therapies and Ebola vaccines will likely be difficult in the under-developed areas most susceptible to Ebola outbreaks. Finally, viral mutations may necessitate the rapid development of new therapeutic agents. Thus, a rapidly engineerable and deployable platform for the portable and scalable production of multiple anti-infectious therapies would be useful for addressing the current Ebola crisis as well as future infectious outbreaks. In this project P. pastoris strains will be designed to produce anti-Ebola mAbs. P. pastoris can be engineered with human glycosylation pathways, can be lyophilized, and is highly efficient at secreting biologics and mAbs, thus enabling industrial-scale production and simplifying purification. The group has already modified P. pastoris with synthetic-biology tools to achieve rapid and specific engineering of strains that manufacture biologic drugs. Distributed and portable production of anti-Ebola mAbs will be achieved by integrating engineered P. pastoris strains with portable micro-bioreactors. The technology also enables new therapeutic molecules to be rapidly generated and scaled-up for testing and deployment against evolving viruses and emerging infections using synthetic biology tools and a highly adaptable biomanufacturing platform.
