Eukaryotic cell display systems
View Patent ↗The present invention provides expression vectors and helper display vectors which can be used in various combinations as vector sets for display of polypeptides on the outer surface of eukaryotic host cells. The expression vector of the invention can be used alone for soluble expression without having to change or reengineer the display vectors. The display systems of the invention are particularly useful for displaying a genetically diverse repertoire or library of polypeptides on the surface of yeast cells, and mammalian cells.
1. A method for displaying a repertoire of polypeptide sequences of interest on yeast host cells, comprising:
a) introducing at least one expression vector comprising an expression cassette encoding a polypeptide of interest fused in frame to a first adapter sequence, into yeast host cells
b) introducing a helper vector encoding a fusion protein consisting of a second adapter sequence fused to an outer surface anchoring protein expressed by the host cells of (a), and
c) maintaining the host cells under suitable conditions for expression of the proteins encoded by the expression cassette and helper vector wherein the polypeptides of interest are displayed on the surface of the yeast host cell wherein the expression vector is selected from pMAT9, and pMAT12, and the helper vector is selected from pMAT7, and pMAT8.
2. The method of claim 1 wherein the repertoire of polypeptide sequences of interest is an antibody scFv library.
3. The method of claim 1 wherein the yeast cells are selected from a group consisting of S. cerevisiae, P. pichia, H. polymorpha , and C. albicans.
4. The method of claim 1 wherein the yeast outer surface anchoring sequences are selected from the group consisting of Aga1 and Aga2, Cwp1, Cwp2, Gaslp, Yap3p, Flo1p, Crh2p, Pir1, Pir2, Pir4, Icwp in S. cerevisiae ; HpSEDI, HpGASI, HpTIPI, HPWPI in H. polymorpha , and Hwp1p, Als3p, Rbt5p in C. albicans.
5. The method of claim 1 wherein the first adapter and second adapter are homodimer sequences.
6. The method of claim 1 wherein the first adapter sequence consists of SEQ ID NO:1 and the second adapter sequence consists of SEQ ID NO: 2.