Patent ID: 9260712
Filing Date: 2016-02-16
CPC Classification: C07K,C12N,G01N

Claim Text:
1. A method of producing yeast host cells that produce immunoglobulins having VH domains, that are fused to Fc domains, and VL domain that are fused to constant regions, and having antigen binding sites with binding specificity for antigens of interest, wherein N-glycans produced by the host cells comprise the structure Man 5 GlcNAc 2 , the method comprising: (a) providing a library of yeast host cells, wherein N-glycans produced by the host cells comprise the structure Man wherein the VH domains are fused to Fc domains; wherein the VL domains are fused to constant regions; wherein the VH domains of the genetically diverse population of immunoglobulins are biased for one or more VH gene families; wherein expression of each VH domain, which is fused to Fc domains, and each VL domain, which is fused to a constant region, of the immunoglobulins, is effected by a second regulatable promoter; wherein each VH domain, that is fused to an Fc, is in an expression cassette, operably linked to a second regulatable promoter and each VL domain, that is fused to a constant region, is in a separate expression cassette, operably linked to a separate second regulatable promoter; and, wherein the yeast host cells display on their surface immunoglobulins comprising VH domains, fused to Fc domains, and VL domains, fused to a constant regions, wherein the library is created by: (b) inducing expression of the first regulatable promoter in the host cells for a time sufficient to produce the capture moieties on the surface of the host cells; wherein the first regulatable promoter is inducible without inducing expression of the second regulatable promoter and the second regulatable promoter is inducible without inducing the expression of the first regulatable promoter; and (c) inhibiting expression of the first regulatable promoter and inducing expression of the second regulatable promoters in the host cells, whereby host cells display immunoglobulins at the surfaces thereof having VH domains, that are fused to Fc domains, and VL domains, that are fused to constant regions, and having the antigen binding sites with binding specificity for the antigen of interest.