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. An apparatus comprising a moldable slab reversibly sealed to a substrate surface, said moldable slab comprising an array of microwells, wherein each microwell has a side or diameter of less than 100 microns and contains not more than a single or a few cell(s), wherein at least one of said microwells contains 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 each said microwell containing the single or few antibody-producing cell(s); wherein the surface of the substrate facing each said microwell containing said single or few antibody-producing cell(s) is bound to monoclonal antibody secreted from said antibody-producing cell(s) in their respective microwell.
2. The apparatus of claim 1 wherein each microwell has a diameter of 10 to 100 microns.
3. The apparatus of claim 2 wherein each microwell has a diameter of 50 to 100 microns.
4. The apparatus of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 10 picoliters to 10 nanoliters.
5. The apparatus of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 1 nanoliter or less.
6. The apparatus of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter.
7. The apparatus of claim 1 wherein the substrate is a glass substrate.
8. The apparatus of claim 1 wherein the moldable slab is made of poly(dimethylsiloxane).
9. The apparatus of claim 1 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure, which protein is bound to said monoclonal antibody secreted from said antibody-producing cell(s).
10. The apparatus of claim 9 wherein the protein is Protein A.
11. The apparatus of claim 9 wherein the protein is Protein G.
12. The apparatus of claim 9 wherein the protein is a secondary antibody.
13. The apparatus of claim 1 wherein the moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.
14. The apparatus of claim 1 wherein each microwell has a diameter of 10 to 100 microns; and the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of nanoliter or less.
15. The apparatus of claim 14 the moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.
16. The apparatus of claim 15 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter.
17. The apparatus 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 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 moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.
18. The apparatus of claim 1 wherein the substrate comprises a plastic material.
19. The apparatus of claim 1 wherein the substrate is substantially planar.
20. The apparatus of claim 15 wherein the surface of the substrate facing said microwells is substantially planar.