Abstract
Recombinant bovine pancreatic aprotinin was produced in Nicotiana species (N. benthamiana and N. excelsiana), a transient expression vector derived from the tobacco mosaic virus genome. Animal-source-free recombinant aprotinin was made in plants grown under controlled (greenhouse) conditions and in the field. Product purified from both production environments showed virtually identical performance and specifications as the bovine-derived pharmaceutical product. Further, the product has excellent stability characteristics and is currently being marketed as a cell culture excipient. Issues, such as oxidative variation of aprotinin product, were addressed by changing growth conditions and by substitution of amino acids for the site of oxidation. The resulting aprotinin variants showed no oxidation, yet retained full activity and production yields. The production parameters of aprotinin provide an affordable way to make high-quality pharmaceuticals in plants. The speed and flexibility of transient expression also allows rapid tailoring of products to address biochemical challenges and product requirements.
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Acknowledgements
We appreciate the efforts of Steve Garger, Terri Cameron, Steve Hume, Kathy Hanley, Josh Morton, and Jennifer Poole in the production and characterization of the r-aprotinin product.
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Pogue, G.P., Vojdani, F., Palmer, K.E., White, E., Haydon, H., Bratcher, B. (2014). Production of Pharmaceutical Grade Recombinant Native Aprotinin and Non-oxidized Aprotinin Variants Under Greenhouse and Field Conditions. In: Howard, J., Hood, E. (eds) Commercial Plant-Produced Recombinant Protein Products. Biotechnology in Agriculture and Forestry, vol 68. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-43836-7_5
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DOI: https://doi.org/10.1007/978-3-662-43836-7_5
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