Addressable antibody arrays and methods of use
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. A kit, the kit comprising:
a plurality of tagged antigens specific for an autoantibody, wherein the autoantibody is associated with a particular disease or disorder, wherein the plurality of tagged antigens transforms the autoantibody into an immunological pair, wherein said tagged antigen further comprises a first support binding member having affinity for an addressable support comprising a second binding member; and
a solid support having a binding member, wherein the binding member is specific for the immunological pair.
2. The kit of claim 1 , wherein said binding member is a protein which binds to said immunological pair.
3. The kit of claim 1 , wherein the 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 kit of claim 1 , wherein the solid support is selected from the group consisting of polystyrene, cellulose, nitrocellulose, a glass bead, and a magnetic bead.
5. The kit of claim 4 , wherein the solid support is a magnetic bead.
6. The kit 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.
7. The kit of claim 1 , wherein said tagged antigen is a plurality of antigens on a support.
8. The kit of claim 1 , wherein said support is a hydrophilic molecule.
9. The kit of claim 8 , wherein said hydrophilic molecule is a polymer.
10. The kit of claim 9 , 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.
11. The kit of claim 1 , wherein said plurality of antigens is attached to said polymer in a “quick-attach” system.
12. The kit of claim 1 , wherein said autoantibody is a plurality of autoantibodies.
13. The kit of claim 12 , wherein said plurality of antigens is specific for said plurality of autoantibodies which form a plurality of immunological pairs.
14. The kit of claim 13 , wherein the increase in affinity from the formation of said plurality of immunological pairs allows for increased detection of a single immunological pair.
15. The kit of claim 1 , wherein said first binding member forms a bound pair with said second binding member on said addressable support.
16. The kit of claim 1 , wherein the kit is used to perform an assay method for detecting an autoantibody in a sample from a subject, wherein the method comprises:
(a) contacting the tagged antigen with a sample having an autoantibody specific for said tagged antigen to transform said autoantibody into an immunological pair;
(b) contacting said immunological pair with the 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.