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 . A kit for use in 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. 
     
     
         2 . The kit of  claim 1 , wherein the disulfated heparin disaccharide is a glucosamine-uronic acid heparin disaccharide. 
     
     
         3 . The kit of  claim 1 , wherein the disulfated heparin disaccharide is IdoAGIcNS-6S. 
     
     
         4 . The kit of  claim 1 , further comprising a solid phase, wherein the disulfated heparin disaccharide is covalently bound to the solid phase. 
     
     
         5 . The kit of  claim 4 , wherein the solid phase is selected from a cuvette, beads, a macrowell plate, a microtiter plate, and a microwell ELISA plate. 
     
     
         6 . The kit of  claim 1 , further comprising a GM1 ganglioside and a second detection antibody. 
     
     
         7 . The kit of  claim 5 , wherein the GM1 ganglioside and the disulfated heparin disaccharide are bound to a solid phase. 
     
     
         8 . The kit of  claim 7 , wherein the solid phase is selected from a cuvette, beads, a macrowell plate, a microtiter plate, and a microwell ELISA plate. 
     
     
         9 . The kit of  claim 1 , wherein the detectable label is an enzyme, radioactive molecule, or a fluorescent agent. 
     
     
         10 . A method of diagnosing a motor neuropathy in a subject, comprising: (a) incubating a test sample from the subject with disulfated heparin disaccharide immobilized on a solid phase, (b) contacting the test sample and disulfated heparin disaccharide with a detection antibody, wherein the detection antibody is detectably labeled and binds to human antibody, and (c) detecting the titer level of the detection antibody indirectly bound to the solid phase, wherein a 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, wherein the detection antibody binds to human IgM antibodies. 
     
     
         11 . 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. 
     
     
         12 . The method of  claim 11 , wherein the disulfated heparin disaccharide is covalently bound to the solid phase. 
     
     
         13 . The kit of  claim 12 , wherein the solid phase is selected from a cuvette, beads, a macrowell plate, a microtiter plate, and a microwell ELISA plate. 
     
     
         13 . The method of  claim 11 , further comprising (c) detecting the titer level of the detection antibody indirectly bound to the disulfated heparin disaccharide. 
     
     
         14 . The method of  claim 13 , wherein a 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. 
     
     
         15 . The method of  claim 11 , wherein the disulfated heparin disaccharide is a glucosamine-uronic acid heparin disaccharide. 
     
     
         16 . The method of  claim 11 , wherein the disulfated heparin disaccharide is IdoA-GIcNS-6S. 
     
     
         17 . The method of  claim 14 , wherein the positive control reference titer is at least about 7,000. 
     
     
         18 . The method of  claim 11 , wherein the detection antibody is detectably labeled with an enzyme, radioactive molecule, or fluorescent agent.