This proposal concerns a low molecular weight protein whose identity we have to conceal due to reasons related to intellectual property rights and patenting. We will call it a Novel Tag (NT) because as you will see below, this protein has a tremendous potential for both academic and industrial research as a protein tag with novel properties. We have discovered that a fraction of His6-tagged NT protein binds to immobilized Ni-nitrilotriacetic acid (Ni-NTA) with extremely high affinity and specificity. Therefore, this project’s aim is to establish a Novel Tag (NT) protein as an attractive alternative to currently used His6-tag. The primary goal of this project is to construct an expression vector and to optimize protocols that would enable the expression, purification and bioanalytical characterization of a set of recombinant proteins fused with NT. This project is conceptualized around the idea that recombinant proteins should be obtained using technologies and materials that are already very widely used (such as the use of FPLC and Ni-NTAcoated beads, chips and plates). However, substantial increase in binding strength and specificity of interaction of NTtagged proteins with Ni-NTA matrices should greatly improve the purity of recombinant proteins as well as the accuracy and sensitivity of bioanalytical methods.
|Effective start/end date||9/1/17 → 9/1/20|