(127a) Transient Recombinant Protein Production in Glycoengineered Plant Cell Suspension Cultures for Rapid Response Applications
AIChE Annual Meeting
Monday, October 29, 2018 - 12:30pm to 12:48pm
As a model protein, we have successfully produced a recombinant anthrax toxin receptor-Fc fusion protein (a fusion between the extracellular domain of the human capillary morphogenesis receptor protein that binds anthrax protective antigen and the Fc domain of human IgG, referred to as CMG2-Fc) by adding Agrobacterium to plant cells in suspension culture. Initial process development studies will be discussed, which resulted in expression levels up to 10 Î¼g CMG2-Fc per g plant cell fresh weight after 7 days of co-culture. Although plants are capable of a wide variety of post-translational modifications, there are slight differences between plant and mammalian glycosylation. To reduce plant-specific glycosylation patterns, Î²(1,2)-xylosyltransferase and Î±(1,3)-fucosyltransferase knockdown glycoengineered host N. benthamiana cell suspension cultures were generated. CMG2-Fc produced by co-culturing Agrobacterium with these glycoengineered N. benthamiana cell suspension cultures resulted in a dramatic reduction in plant-specific N-glycans compared to CMG2-Fc produced in wild type N. benthamiana plants. In addition to mitigating the risk of an immune response to plant-specific glycans, this proof of concept data demonstrates a method that could be further modified to enhance a productâs efficacy and stability by tuning its N-glycan distribution. Finally, data from techno-economic analysis of both upstream and downstream unit operations performed using SuperPro Designer software will be presented. These simulation results will guide process development work by identifying yield and recovery levels required, as well as key cost drivers, for cost-effective, large-scale production using this novel platform.