Screening assays and methods
Screening assays and methods of performing such assays are provided. In certain examples, the assays and methods may be designed to determine whether or not two or more species can associate with each other. In some examples, the assays and methods may be used to determine if a known antigen binds to an unknown monoclonal antibody.
1. A method of producing a microarray of monoclonal antibodies comprising:
(a) reversibly sealing a moldable slab to a substrate, said moldable slab comprising an array of microwells, each microwell having a side or diameter of less than 100 microns and containing not more than a single or a few cell(s), at least one of said microwells containing the single or few cell(s), wherein the cell(s) are antibody-producing cell(s) that secrete monoclonal antibodies in a volume of 10 nanoliters or less of fluid in said microwell; wherein a surface of the substrate facing said microwells is modified such that it acts to retain monoclonal antibody on the surface of the substrate; and
(b) allowing said antibody-producing cell(s) to secrete said monoclonal antibodies in said volume such that monoclonal antibody secreted from the antibody-producing cell(s) in said volume is retained in a detectable amount on the surface of the substrate sealed to the moldable slab.
2. The method of claim 1 wherein each microwell has a diameter of 10 to 100 microns.
3. The method of claim 2 wherein each microwell has a diameter of 50 to 100 microns.
4. The method of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 10 picoliters to 10 nanoliters.
5. The method of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 1 nanoliter or less.
6. The method of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter.
7. The method of claim 1 wherein the detectable amount is an amount suitable to detect association of a labeled species with said monoclonal antibody.
8. The method of claim 7 wherein the labeled species is a fluorescently labeled antigen.
9. The method of claim 1 wherein the substrate is a glass substrate.
10. The method of claim 1 wherein the moldable slab is made of poly(dimethylsiloxane).
11. The method of claim 1 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure.
12. The method of claim 11 wherein the protein is Protein A.
13. The method of claim 11 wherein the protein is Protein G.
14. The method of claim 11 wherein the protein is a secondary antibody.
15. The method of claim 1 wherein the steps of reversibly sealing and allowing are performed in less than 24 hours.
16. The method of claim 15 wherein the steps of reversibly sealing and allowing are performed in less than about 12 hours.
17. The method of claim 1 wherein the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate.
18. The method of claim 1 wherein each microwell has a diameter of 10 to 100 microns; the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 1 nanoliter or less; and the detectable amount is an amount suitable to detect association of a labeled species with said monoclonal antibody.
19. The method of claim 18 wherein the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate.
20. The method of claim 19 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter.
21. The method of claim 1 wherein each microwell has a diameter of 10 to 100 microns; the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter; the detectable amount is an amount suitable to detect association of a fluorescently labeled antigen with said monoclonal antibody; the substrate is a glass substrate; the moldable slab is made of poly(dimethylsiloxane); the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure; and the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate.
22. The method of claim 1 wherein the substrate is comprised of a plastic material.
23. The method of claim 1 wherein the surface facing said microwells is substantially planar.