Nanobody Discovery Platform and Related Methods

A nucleic acid comprising one or more inducible gene for expressing in a yeast cell:

• a first fusion protein comprising amino acid sequences of:
• an anchor peptide selected for anchoring the first fusion protein in a cell wall of the cell, a nanobody, and an scFv (FAP) for binding a first fluorophore;
• and
• a second fusion protein comprising amino acid sequences of a secretion signal peptide and an scFv (FAP) for binding a second fluorophore,
• wherein the first fluorophore increases fluorescent emissions, e.g., at least 1000-fold, at an excitation wavelength when bound by the scFv of the first fusion protein, the second fluorophore increases fluorescent emissions, e.g., at least 1000-fold, at an excitation wavelength when bound by the scFv of the second fusion protein, and the fluorescent emission of the scFv-bound first fluorophore is detectably different from that of the scFv-bound second fluorophore, wherein the nucleic acid further comprises a barcode for identifying a target protein domain (TPD) in the coding sequence of the first fusion protein and wherein the first fusion peptide optionally comprises one or more type IIS restriction endonuclease recognition site-terminated DNA modules.