This I-CORPS proposal addresses the following statement: Significant effort is associated with the purification of a biological for applications which include biopharmaceutical development. Creating a series of custom strains of Escherichia coli that express minimized sets of nuisance proteins will be of commercial value, with their creation uniquely based on knowledge of the separatome of E. coli. Producing recombinant products, therapeutics in particular, occur with and without the use of an affinity tail to dictate purification. In its absence, production with E. coli can be a challenge: bioseparation steps are developed somewhat arbitrarily; the presence of host cell proteins reduces separation step efficiency; and the tradeoff between overall yield and purity may not be best. Contrastingly, the use of an affinity tail helps reduce the chromatographic space but still can be plagued by: co-adsorbing / co-eluting proteins; removal of the affinity tail; and ligand cost.
By developing E. coli strains that display a minimal contaminant pool, it is believed that industrial-relevant proteins can be developed more cost-effectively. The current effort will prioritize subsequent development based on potential customer feedback collected as a result of I-Corps support. If a strong business model can be discovered, the team plans to pursue this activity in a newly-formed venture.
Intellectual Merit One of the bottlenecks associated with the manufacture of commodity enzymes like those used in high fructose corn syrup production, and therapeutic proteins like those found in an insulin shot, is the ability to efficiently purify the target from a complicated mixture. The I-Corps grant equiped our team to validate this premise regarding manufacturing and to align our fundamental work in the areas of metabolic engineering and bioseparation with the needs of industry. During the grant period and beyond, we visited with "captains of industry" - persons with domain expertise in the area of enzyme / therapeutic manufacturing, and determined that the cell lines we are developing address this bottleneck. Moreover, the methodology and data are of sufficient practical benefit to also offer "blueprints" of efficient cell lines that increase bioseparation efficiency as additional products. Broader Impact This project positively impacts enzyme / therapeutic manufacturing that increases throughput for a broad spectrum of end products. For example, more product can be purified each time an [ion exchange] chromatography column is cycled through loading, wash, and product elution. Boston Mountain Biotech, LLC, a WOSB, was created as a result of the I-Corps Program and strives to bring these types of employment opportunities to the State of Arkansas.
