High affinity VEGF-receptor antagonists
View Patent ↗A cell-based screen is reported can be used to identify specific receptor-binding compounds in a combinatorial library of peptoids (N-alkylglycine oligomers) displayed on beads. This strategy was applied to the isolation of Vascular Endothelial Growth Factor Receptor 2 (VEGFR2)-binding peptoids, which were optimized to create lead compounds with high affinity for VEGFR2. One of these peptoids was shown to be an antagonist of VEGF-VEGFR2 interaction and receptor function.
1. A method of screening a library of peptoid ligands for binding specificity to a cell population mixture comprising:
(a) providing a peptoid ligand library;
(b) providing a mixture of cells, wherein said mixture comprises a first cell population of a particular cell type which presents a target cell surface structure and a second cell population of the same or different cell type which does not present the target cell surface structure and wherein the first cell population and the second cell population are differentially labeled with distinct detectable quantum dots;
(c) contacting said peptoid ligand library with the mixture of cells; and
(d) determining the binding specificity of the peptoid ligand library by detecting the differentially labeled cell populations bound to the peptoid ligands of the library.
2. The method of claim 1 , wherein the peptoid ligands of the library are displayed on a support.
3. The method of claim 1 , wherein the cell type of said first cell population is selected from the group comprising a T-cell, a B-cell, an epithelial cell, an endothelial cell, a kidney cell, a cancer cell, a pathogen-infected cell or any other culturable cell type.
4. The method of claim 1 , wherein the cell type of said first cell population is a single-celled organism.
5. The method of claim 1 , wherein the target cell surface structure comprises an antibody or T-cell receptor, a growth factor receptor, a cell matrix protein, a lectin, a carbohydrate, a lipid or a cell surface antigen.
6. The method of claim 1 , wherein at least one of said first and second cell populations is genetically modified.
7. The method of claim 1 , wherein the differential label comprises a colorimetric, fluorometric, bioluminescent or chemilluminescent label.
8. The method of claim 2 , further comprising identifying the structure or identity of one or more peptoid ligands bound to the differentially labeled cell populations.
9. The method of claim 1 , wherein said target cell surface structure is a VEGF2 receptor.
10. The method of claim 1 , wherein said target cell surface structure is an antigen.
11. The method of claim 1 , wherein said target cell surface structure is a lipid.
12. The method of claim 1 , wherein said target cell surface structure is a carbohydrate.
13. The method of claim 1 , wherein said quantum dots are internalized into the cells.
14. The method of claim 1 , wherein said first cell population and second cell population are B-cell or T-cell types.
15. The method of claim 1 , wherein the cell type of said first cell population is a cancer cell.
16. The method of claim 1 , wherein the cell type of said first cell population is a pathogen cell or pathogen-infected cell.
17. The method of claim 1 , wherein the cell types of said first and second cell populations differ by a specific mutation in a particular gene.
18. The method according to claim 1 , wherein the cell type of said first cell population is identical to the cell type of said second cell population.
19. The method of claim 1 , wherein said peptoid ligand library has greater than 300,000 distinct oligomers and has at least one monomer selected from the group consisting of: