Optimization of Therapeutic Protein Production in an Aquatic Plant Expression System Using DOE
Lynn Dickey, PhD, Vice President of Research and Technology Development; Vincent Wingate, Associate Director of Upstream Process Development and Plant Biology – Biolex Therapeutics
Biolex Therapeutics uses a novel biomanufacturing platform (LEX SystemSM) based on the aquatic higher plant Lemna minor, genetically engineered to produce recombinant biopharmaceutical proteins. We use a small-scale process model to optimize critical factors to maximize the yield and quality of recombinant biopharmaceutical proteins. Using the design of experiments (DOE) capabilities in JMP, we will discuss what information is required prior to setting up a DOE, how to set up a custom DOE, and how to analyze and visualize the data. We will exemplify the JMP DOE platform’s powerful design, analytical and graphic capabilities that have allowed us to optimize critical parameters and identify critical parameter interactions and optimal ranges (both for environmental and media components) that affect the yield and quality of several different therapeutic proteins. A designed experiment will be presented that optimizes carbon dioxide levels, nitrogen concentration and environmental factors to maximize the yield of a therapeutic protein. It is well known that two of the major nutritional components required for increases of plant biomass are higher levels of nitrogen and carbon dioxide. It is not yet fully defined in the LEX SystemSM what combination of nitrogen, carbon dioxide and environmental factors (e.g., light intensity and temperature) are required for maximal expression of a recombinant protein. Using our small-scale process model, we will show how we have used custom DOE experiments with these critical parameters to optimize the expression of a vaccine antigen recombinant protein.