Antibodies to disulfated heparin disaccharide in the diagnosis of neuropathy

Abstract

Methods aiding in the diagnosis of certain neuropathies are disclosed, in which the titer of antibodies to a disulfated heparin disaccharide is assessed in a test sample from a subject. Also disclosed are apparatus and kits that can be used in the methods of the invention.

1 .- 18 . (canceled) 
     
     
         19 . A kit for diagnosing a neuropathy in a subject, comprising an isolated or synthesized disulfated heparin disaccharide and a detection antibody that specifically binds to human IgM, wherein the detection antibody comprises a detectable label. 
     
     
         20 . The kit of  claim 19 , wherein the disulfated heparin disaccharide is a glucosamine-uronic acid heparin disaccharide. 
     
     
         21 . The kit of  claim 19 , wherein the disulfated heparin disaccharide is IdoAGlcNS-6S. 
     
     
         22 . The kit of  claim 19 , further comprising a solid phase, wherein the disulfated heparin disaccharide is covalently bound to the solid phase. 
     
     
         23 . The kit of  claim 22 , wherein the solid phase is selected from a cuvette, beads, a macrowell plate, a microtiter plate, and a microwell ELISA plate. 
     
     
         24 . The kit of  claim 19 , further comprising a GM1 ganglioside and a second detection antibody. 
     
     
         25 . The kit of  claim 23 , wherein the GM1 ganglioside and the disulfated heparin disaccharide are bound to a solid phase. 
     
     
         26 . The kit of  claim 25 , wherein the solid phase is selected from a cuvette, beads, a macrowell plate, a microtiter plate, and a microwell ELISA plate. 
     
     
         27 . The kit of  claim 19 , wherein the detectable label is an enzyme, radioactive molecule, or a fluorescent agent. 
     
     
         28 . A method of preparing a sample from a subject suspected of suffering from a motor neuropathy, comprising: (a) incubating a test sample from the subject with disulfated heparin disaccharide, and (b) contacting the test sample and disulfated heparin disaccharide with a detection antibody, wherein the detection antibody is detectably labeled and binds to human IgM antibodies. 
     
     
         29 . The method of  claim 28 , wherein the disulfated heparin disaccharide is covalently bound to the solid phase. 
     
     
         30 . The method of  claim 29 , wherein the solid phase is selected from a cuvette, beads, a macrowell plate, a microtiter plate, and a microwell ELISA plate. 
     
     
         31 . The method of  claim 28 , further comprising (c) detecting a titer level of the detection antibody indirectly bound to the disulfated heparin disaccharide. 
     
     
         32 . The method of  claim 31 , wherein the titer level of detection antibody equal to or greater than a positive control reference titer is indicative of the presence of a motor neuropathy in the subject. 
     
     
         33 . The method of  claim 28 , wherein the disulfated heparin disaccharide is a glucosamine-uronic acid heparin disaccharide. 
     
     
         34 . The method of  claim 28 , wherein the disulfated heparin disaccharide is IdoA-GlcNS-6S. 
     
     
         35 . The method of  claim 32 , wherein the positive control reference titer is at least about 7,000. 
     
     
         36 . The method of  claim 28 , wherein the detection antibody is detectably labeled with an enzyme, radioactive molecule, or fluorescent agent. 
     
     
         37 . A detection antibody that specifically binds a human antibody that specifically binds to an isolated or synthesized disulfated heparin disaccharide, wherein the detection antibody comprises a detectable label, wherein the human antibody is a human IgM antibody, and wherein the titer level of the detection antibody is indicative of the levels of autoantibody against disulfated heparin disaccharide in a test sample from the subject. 
     
     
         38 . The detection antibody of  claim 37 , wherein the disulfated heparin disaccharide is (a) a glucosamine-uronic acid heparin disaccharide, or (b) IdoA-GlcNS-6S.