Surface anchored light chain bait antibody display system
The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. Embodiments of the invention provide a system in which a bait complexed with a monovalent antibody fragment can be captured prior to secretion in a host cell by virtue of surface displaying an antibody light chain and utilizing the covalent interaction of this light chain with the heavy chain of an antibody molecule that is co-expressed in the same host. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of using the system for identifying antibodies that bind specifically to an antigen of interest.
1. An isolated bait polypeptide comprising a full-length immunoglobulin light chain fused, optionally by a peptide linker, to a surface anchor polypeptide or a functional fragment thereof which bait polypeptide is optionally amino-terminally fused to a signal peptide; wherein the surface anchor polypeptide is SED-1 or a functional fragment thereof.
2. The polypeptide of claim 1 wherein the immunoglobulin light chain comprises a kappa or lambda constant immunoglobulin domain.
3. An isolated polynucleotide encoding the polypeptide of claim 1 .
4. A vector comprising the polynucleotide of claim 3 .
5. An isolated host cell comprising the polypeptide of claim 1 or a polynucleotide encoding said polypeptide, wherein the host cell is a Pichia or Saccharomyces cerevisiae host cell.
6. The host cell of claim 5 further comprising one or more polynucleotides encoding full-length immunoglobulin light chain; and/or one or more polynucleotides encoding full-length immunoglobulin heavy chain.
7. The host cell of claim 5 , wherein the Pichia cell is a Pichia pastoris cell.
8. The host cell of claim 6 wherein the bait polypeptide is located on the surface of the host cell membrane.
9. A composition comprising the host cell of claim 6 , further comprising a non-tethered full antibody comprising said full-length immunoglobulin light and heavy chains; optionally, complexed with an antigen.