Surface, anchored FC-bait antibody display system
The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. A bait complexed with a monovalent antibody fragment can be expressed on the surface of the host cell wherein the fragment may be assayed for antigen binding while full antibody is simultaneously secreted from the host cell. Methods of using the system for identifying antibodies that bind specifically to an antigen of interest are also provided. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of use thereof.
1. A method for making an antibody comprising culturing an isolated eukaryotic host cell in a growth medium under conditions allowing expression of an immunoglobulin light chain and an immunoglobulin heavy chain of said antibody, wherein the host cell is selected from the group consisting of a yeast and filamentous fungi; wherein the eukaryotic host cell comprises:
(i) a first polynucleotide encoding said immunoglobulin light chain; and a second polynucleotide encoding said immunoglobulin heavy chain wherein the heavy chain is VH—CH1-CH2-CH3, each of said first and second polynucleotides operably associated with a promoter for expressing the immunoglobulin heavy and light chains; and
(ii) a third polynucleotide encoding a bait comprising a heavy Fc immunoglobulin domain fused to a surface anchor polypeptide on the surface of said eukaryotic host cell, operably associated with a regulatable promoter for expressing the polynucleotide encoding the bait;
wherein the Fc of said bait when expressed forms a complex with the Fc of the immunoglobulin heavy chain, which complex is displayed on the surface of the host cell; and
wherein when expression of the bait is inhibited, said antibody is secreted from said eukaryotic host cell.
2. The method of claim 1 , wherein the yeast is Pichia.
3. The method of claim 2 , wherein the Pichia is Pichia pastoris.
4. The method of claim 1 , wherein the yeast is S. cerevisiae.
5. The method of claim 1 , wherein said antibody is isolated from said eukaryotic host cell and medium.