UNS:Thermal control of avidity for separation of biologicals

North Carolina State University, Raleigh NC

Investigators

Abstract

1511227 Rao North Carolina State U Affinity chromatography exploits the highly specific interaction between molecules or ligands immobilized on a surface and the target species of interest. In the proposed work affinity adsorbents will be generated with the proposed molecules or ligands for both small and large scale separations using the same synthesis scheme. These affinity adsorbents are expected to be robust and amenable to multiple re-uses after sterilization. Yet, it is expected that production of adsorbents for single-use applications with high value biologicals will still be cost-effective. Collaboration with a North Carolina State-based company will be undertaken to explore the development of commercial membrane-based separation systems for complex biologicals. To identify temperature-sensitive affinity ligands, a peptide-based scaffold that changes conformation in response to changes in temperature will be evaluated. Affinity ligands for two model targets will be generated: CD117, which is expressed on hematopoietic stem cells, and TOM22, a mitochondrial protein. Subsequently, the effectiveness of these ligands will be evaluated in the context of cell and mitochondrial separation. Accordingly, combinatorial libraries of peptides will be screened to identify peptides binders for the model targets. Thermo-responsive ligands will also be constructed using the peptides generated. Subsequently, the effect of temperature on the binding affinity of these ligands will be evaluated. Finally, the use of magnetic beads functionalized with the selected ligands will be evaluated in the context of separation of CD117+ cells and mitochondria.