PFI: TT: Development of 5-Minute Protein Analyses

The broader impact/commercial potential of this Partnerships for Innovation - Technology Translation (PFI-TT) project includes development of a convenient device that can determine protein concentrations in 5 minutes. Current methods for protein analysis often employ expensive kits that require more than 1 hour to obtain results. The new technology will enable regular monitoring of protein concentrations during their manufacturing and processing.

The markets for some individual antibodies as well as other proteins are in the range of $10 billion per year, and these biomolecules are vital in treating diseases such as cancer, diabetes, and anemia. Improving the production of these proteins by monitoring their expression and processing is an important step in enhancing their availability. The research may yield a simple platform for protein analysis in multi-well plates used in most laboratories.

Given the size of the therapeutic protein market (>$250 billion/year), tools for analysis of therapeutic protein production should have a market of many million dollars per year.

This project will use flow through functionalized microporous membranes in 96-well plates to enable capture and detection of proteins in minutes. In contrast, typical protein analysis platforms rely on passive diffusion to bring proteins to binding sites; Analysis times are >1 h. The research includes a method for functionalizing the porous membranes with peptides that bind specifically to a given protein.

The high density of peptide immobilization and the high surface area to volume ratio in the membranes allows capture of large amounts of protein in a short time to achieve rapid, sensitive quantitation. The research aims to demonstrate a general method for the analyses of antibodies in cell-culture broths as well as comparable analyses times, detection limits, percent recoveries, and precision when compared to commercial immunosorbent assays.

Additionally, the work may establish membrane-based capture and analysis of other important proteins such as insulin and erythropoietin. Success in these studies may demonstrate the promise of functionalized membrane plates for regularly monitoring the production and processing of important commercial proteins. The research will also demonstrate the advantages of this method over current techniques.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.