The goal of this project is to develop bioprocess-compatible, second generation nanoCLAMP affinity resins to several high-value plasma-based therapeutics and characterize their performance metrics to provide tools for improving process efficiency. The plasma-derived therapy market ($19B) is expected to double by 2033, with new target proteins being rapidly developed. Due to the high cost of the raw material for these therapeutics there is a critical need for efficient recovery of the proteins in plasma. A bioprocess-compatible affinity chromatography platform capable of high percentage and specificity recovery of plasma proteins would enable lower cost production of existing therapeutics and drive development of new ones. The incorporation of affinity chromatography into downstream processing of non-tagged proteins is hampered by a lack of bioprocess- compatible affinity ligands with adequate affinity, specificity, and durability. Most available protein-based affinity ligands are not caustic-stable, limiting their use in repeated cycles of cleaning in place. This work seeks to apply a novel bioprocess-compatible antibody mimetic platform, called the nanoCLAMP, to affinity chromatography processing of human plasma to streamline downstream purification of plasma-based therapeutics. The specific aims of this proposal are to: 1. Identify panels of nanoCLAMPs to 5 industrially relevant plasma-based therapeutics 2. Select leads with desired performance metrics. 3. Purify targets from plasma using lead resins and optimize linker lengths and resin composition to maximize % recovery and purity. Scale up production of lead affinity chromatography resins and determine performance metrics of final products (10 ml scale) for purification of targets from plasma. The successful completion of this project will yield a versatile platform for the development of affinity chromatography resins utilizing the innovative nanoCLAMP ligand, allowing for the high-purity isolation of protein components from plasma. The customizable nature of nanoCLAMP ligands will enable tailoring the resins for specific blood components, addressing current challenges in the purification of complex biological samples. The outcomes of this research have the potential to revolutionize protein purification processes in biopharmaceutical and clinical applications, enhancing the efficiency and reliability of downstream processing.
Public Health Relevance Statement: The human plasma-based therapeutics market is responsible for therapies that treat a wide range of human diseases, such as idiopathic thrombocytopenic purpura, von Willebrand disease, hemophelia, and primary immunodeficiency, and many others. Because the raw material for these therapeutics is expensive and precious, downstream processing improvements are necessary to make existing therapeutics more cost effective and to drive discovery of new ones. The goal of this project is to apply a novel bioprocess-compatible antibody mimetic platform called the nanoCLAMP for the efficient affinity-purification of human plasma proteins. Terms:
