# US 8,772,049 B2 · Screening assays and methods > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,772,049 B2 - Document ID: US8772049B2 - Status: Granted - Kind: B2 - Application number: 13/960,757 - Filed: 2013-08-06 - Published: 2013-12-19 - Granted: 2014-07-08 - Earliest claimed: 2005-09-16 - Assignees: President and Fellows of Harvard College - Applicants: President and Fellows of Harvard College - Inventors: J. Christopher Love; Hidde L. Ploegh; Jehnna Ronan - Web page: https://patentgrep.com/patent/US8772049B2 ## CPC - B01J 19/0046: {Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays (synthesis methods per se C40B50/00)} - B01L 3/50853: {with covers or lids (closures for test tubes B01L3/50825)} - B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - G01N 33/505: {involving T-cells} - G01N 33/5052: {involving B-cells} - G01N 33/531: Production of immunochemical test materials - G01N 33/543: with an insoluble carrier for immobilising immunochemicals - G01N 33/54366: {Apparatus specially adapted for solid-phase testing} - G01N 33/577: involving monoclonal antibodies {binding reaction mechanisms characterised by the use of monoclonal antibodies (G01N33/5302 - G01N33/576 take precedence)} - G01N 33/6854: {Immunoglobulins} - B01J 2219/00317: Microwell devices, i.e. having large numbers of wells - B01J 2219/00527: Sheets - B01J 2219/00585: Parallel processes - B01J 2219/00675: In-situ synthesis on the substrate - B01J 2219/00722: Nucleotides - B01J 2219/00725: Peptides - B01J 2219/00734: Lipids - B01J 2219/0074: Biological products - B01L 2200/025: Align devices or objects to ensure defined positions relative to each other - B01L 2200/12: Specific details about manufacturing devices - B01L 2300/0819: Microarrays; Biochips - B01L 2300/12: Specific details about materials - Y10S 436/809: Multifield plates or multicontainer arrays - Y10T 436/24: Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS - Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION ## Abstract 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. ## Claims ### Claim 1 A method of screening a monoclonal antibody 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, wherein microwells of said array contain a single or a few antibody-producing cell(s) that secrete monoclonal antibodies in a volume of 10 nanoliters or less of fluid in said microwells; wherein the substrate's surface facing the microwells is modified such that it acts to retain such secreted monoclonal antibodies on the surface; (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 by the surface of the substrate sealed to the moldable slab; (c) removing the substrate from the moldable slab; and (d) exposing the retained monoclonal antibody to an antigen to test the retained monoclonal antibody for a desired binding specificity. ### Claim 2 (depends on 1) The method of claim 1 wherein each microwell has a diameter of 10 to 100 microns. ### Claim 3 (depends on 2) The method of claim 2 wherein each microwell has a diameter of 50 to 100 microns. ### Claim 4 (depends on 1) The method of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 10 picoliters to 10 nanoliters. ### Claim 5 (depends on 1) The method of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 1 nanoliter or less. ### Claim 6 (depends on 1) The method of claim 1 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter. ### Claim 7 (depends on 1) The method of claim 1 wherein the antigen is labeled. ### Claim 8 (depends on 7) The method of claim 7 wherein the antigen is fluorescently labeled. ### Claim 9 (depends on 1) The method of claim 1 wherein the substrate is a glass substrate. ### Claim 10 (depends on 1) The method of claim 1 wherein the moldable slab is made of poly(dimethylsiloxane). ### Claim 11 (depends on 1) 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. ### Claim 12 (depends on 11) The method of claim 11 wherein the protein is Protein A. ### Claim 13 (depends on 11) The method of claim 11 wherein the protein is Protein G. ### Claim 14 (depends on 11) The method of claim 11 wherein the protein is a secondary antibody. ### Claim 15 (depends on 1) The method of claim 1 wherein steps (a)-(d) are performed in less than 24 hours. ### Claim 16 (depends on 15) The method of claim 15 wherein steps (a)-(d) are performed in less than about 12 hours. ### Claim 17 (depends on 1) The method of claim 1 wherein the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate. ### Claim 18 (depends on 1) The method 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 1 nanoliter or less. ### Claim 19 (depends on 18) The method of claim 18 wherein the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate. ### Claim 20 (depends on 19) The method of claim 19 wherein the antibody-producing cell(s) secrete the monoclonal antibodies in a volume of 100 picoliters to 1 nanoliter. ### Claim 21 (depends on 1) 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 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. ### Claim 22 (depends on 2) The method of claim 2 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 23 (depends on 3) The method of claim 3 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 24 (depends on 4) The method of claim 4 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 25 (depends on 5) The method of claim 5 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 26 (depends on 8) The method of claim 8 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 27 (depends on 15) The method of claim 15 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 28 (depends on 17) The method of claim 17 wherein the surface of the substrate is coated with a protein that recognizes the constant region of an antibody's structure. ### Claim 29 (depends on 1) The method of claim 1 wherein the substrate comprises a plastic material. ### Claim 30 (depends on 19) The method of claim 19 wherein the surface of the substrate facing said microwells is substantially planar.