Patent ID: 8067339

Claim:
A method for producing yeast host cells that express an immunoglobulin of interest, comprising: (a) providing yeast host cells that include a first nucleic acid molecule operably linked to a GUT1 promoter and which encodes a capture moiety comprising a cell surface anchoring protein fused to protein A; (b) transfecting the yeast host cells with a plurality of nucleic acid molecules encoding a genetically diverse population of heavy and light chains of an immunoglobulin wherein at least one of the heavy or light chain encoding nucleic acid molecules is operably linked to a GAPDH promoter to produce a plurality of genetically diverse yeast host cells capable of displaying immunoglobulins on the surface thereof; (c) growing the yeast host cells in the presence of glycerol for a time sufficient to produce the capture moiety on the surface of the yeast host cells and then growing the yeast cells in a medium that lacks glycerol but contains dextrose to produce yeast host cells in which the immunoglobulins expressed in the yeast host cells are captured by the capture moiety on the surface of the yeast host cells; (d) contacting the host cells with a detection means that specifically binds to the immunoglobulin of interest; and (e) isolating yeast host cells in which the detection means is bound to the immunoglobulin of interest to produce the yeast host cells that express the immunoglobulin of interest.