High-throughput screening of mammalian signal peptide library
2The Russell Berrie Nanotechnology Institute, Technion Israel institute of Technology, Israel
3Chemical Engineering, Guangdong Technion Israel institute of Technology, China
Understanding the relationship between protein and the presence of signal peptides (SPs) is a fundamentally important question in biology and for various biotechnological applications. Although most of the applications using SPs are focused on improving protein production and secretion in prokaryotes and yeast, the emerging fields of "alternative proteins" and "biologics" for the food- and pharmaceutical-sectors are expected to be highly reliant on the production and secretion of proteins from mammalian cells. At present, protein production and secretion are still poorly understood in mammalian systems, and consequently many protein-production applications are either ineffective or sub-optimal at best. Here, we describe a high-throughput screening fluorescent-based method for mammalian secretory SPs. We demonstrate the feasibility of our approach by engineering several functional SPs, and demonstrating successful secretion at differing titers of mCherry. Finally, we discuss future applications based on this technology on problems involving the DKK1 AND Wnt3a pathways.