Yeast cell surface display of proteins and uses thereof
US6331391A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jan 20, 1998 |
| Grant date | Dec 18, 2001 |
| Priority date | — |
| Expiry date | Jan 20, 2018 |
Classification
- Technology area (CPC —)General
Abstract
The present invention provides a genetic method for tethering polypeptides to the yeast cell wall in a form accessible for binding to macromolecules. Combining this method with fluorescence-activated cell sorting provides a means of selecting proteins with increased or decreased affinity for another molecule, altered specificity, or conditional binding. Also provided is a method for genetic fusion of the N terminus of a polypeptide of interest to the C-terminus of the yeast Aga2p cell wall protein. The outer wall of each yeast cell can display approximately 10.sup.4 protein agglutinins. The native agglutinins serve as specific adhesion contacts to fuse yeast cells of opposite mating type during mating. In effect, yeast has evolved a platform for protein-protein binding without steric hindrance from cell wall components. As one embodiment, attaching an scFv antibody fragment to the Aga2p agglutinin effectively mimics the cell surface display of antibodies by B cells in the immune system for affinity maturation in vivo. As another embodiment, T cell receptor mutants can be isolated by this method that are efficiently displayed on the yeast cell surface, providing a means of altering …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.