Addressable antibody arrays and methods of use
View Patent ↗Systems and assay methods are disclosed for detecting an autoantibody in a sample. In certain instances, the systems and methods employ a mass tag releasably connected to an antigen. The tag is thereafter released for detection. A tag can be detected by mass spectrometry or in certain instances the tag is fluorescent. Methods for diagnosing a disease or disorder in a subject are also disclosed.
1. An assay method for detecting an autoantibody in a sample from a subject, said method comprising:
(a) contacting a tagged antigen with a sample having an autoantibody specific for said tagged antigen to transform said autoantibody into an immunological pair, wherein said tagged antigen is a plurality of antigens on a support;
(b) contacting said immunological pair with a solid support having a binding member specific for said immunological pair to transform said immunological pair into a protein complex;
(c) separating said protein complex from said sample to form an isolated protein complex; and
(d) releasing said tag from said isolated protein complex for detection.
2. The assay method of claim 1 , wherein said tag is detected by mass spectrometry.
3. The assay method of claim 1 , wherein said binding member is selected from the group consisting of an antigen, an antibody, biotin, avidin, streptavidin, anti-biotin; folate, folate-binding protein, IgG, Protein A, Protein G, a carbohydrate, lectin, and a nucleic acid.
4. The assay method of claim 1 , wherein said binding member is a protein which binds to said immunological pair.
5. The assay method of claim 1 , wherein the solid support is selected from the group consisting of polystyrene, cellulose, nitrocellulose, a glass bead, and a magnetic bead.
6. The assay method of claim 5 , wherein the solid support is a magnetic bead.
7. The assay method of claim 1 , wherein said sample is a member selected from the group consisting of whole blood, serum, plasma, urine, seminal fluid, and saliva.
8. The assay method of claim 7 , wherein said sample is whole blood.
9. The assay method of claim 1 , wherein said autoantibody is derived from a subject having an autoimmune disease selected from the group consisting of rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, systematic lupus erythematosus, Sjogren's syndrome, ankylosing spondylitis, psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, graft-vs-host disease, and scleroderma.
10. The assay method of claim 1 , wherein said support is a hydrophilic molecule.
11. The assay method of claim 10 , wherein said hydrophilic molecule is a polymer.
12. The assay method of claim 11 , wherein said polymer is a member selected from the group consisting of polyethylene glycol, dextran, dextran carboxylic acid polyvinyl pyrrolidone, sugar alcohols, polyoxyethylene polyoxypropylene glycol, and a mixture thereof.
13. The assay method of claim 11 , wherein said plurality of antigens is attached to said polymer in a “quick-attach” system.
14. The assay method of claim 1 , wherein said autoantibody is a plurality of autoantibodies.
15. The assay method of claim 14 , wherein said plurality of antigens is specific for said plurality of autoantibodies which form a plurality of immunological pairs.
16. The assay method of claim 15 , wherein the increase in affinity from the formation of said plurality of immunological pairs allows for increased detection of a single immunological pair.
17. The assay method of claim 1 , wherein said tagged antigen further comprises a first binding member having affinity to an addressable support comprising a second binding member.
18. The assay method of claim 17 , wherein said first binding member forms a bound pair with said second binding member on said addressable support.
19. A method for diagnosing a disease or disorder in a subject, said method comprising:
(a) contacting a tagged antigen with a sample from said subject having an autoantibody specific for said tagged antigen to transform said tagged antigen into an immunological pair, wherein said tagged antigen is a plurality of antigens on a support;
(b) contacting said immunological pair with a solid support having a binding member specific for said immunological pair to transform said immunological pair into a protein complex;
(c) separating said protein complex from said sample to form an isolated protein complex;
(d) releasing said tag from said isolated protein complex for detection; and
(e) detecting the amount of said tag, wherein the amount of said tag is indicative of the amount of said autoantibody, and wherein said disease or disorder is determined to be present when the amount of said autoantibody differs from a control value representing the amount of said autoantibody in a sample from a subject not having said disease or disorder.