(27bj) Heterologous Derived Proteins Tethered to the Surface of Yeast: Arming Saccharomyces Cerevisiae with Sars-Cov-2 Receptor Binding Domain and Green Fluorescent Protein.

Yeast surface display is a molecular biology technique that arms the exterior of a cell with proteins to provide new functionality. Predominantly, display involves the insertion of individual genes into an expression cassette which normally limits the number of heterologous proteins from one to five. Developing a new yeast-based strategy that permits the multiplexing of numerous proteins on the surface, regardless of source be it yeast, bacterial, or other, is the focus of this study. This presentation will describe our efforts with S. cerevisiae that is armed with proteins recombinantly produced in Escherichia coli. An optimized oxidation buffer and kinetic model of surface uptake will be shown, validated by confocal and fluorescence microscopy and flow cytometry. Coculturing techniques to prepare a one-pot expression will be discussed using batch and novel bioreactor conditions, along with applications for the multiplexed yeast for diagnostic applications.