Assay reagents for a neurogranin diagnostic kit

Abstract

The present invention relates to the field of biomarkers. More specifically, the present invention relates to assay reagents useful in detecting neurogranin. In a specific embodiment, the present invention provides an isolated antibody or fragment thereof that specifically binds to neurogranin. In another embodiment, the present invention provides a polynucleotide aptamer that specifically binds neurogranin.

1 . An isolated antibody or fragment thereof that specifically binds to neurogranin. 
     
     
         2 . An isolated antibody or fragment thereof that specifically binds to amino acids 1-78 of SEQ ID NO:11. 
     
     
         3 . An isolated antibody or fragment thereof that specifically binds to amino acids 55-78 of SEQ ID NO:11. 
     
     
         4 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody or fragment thereof is polyclonal. 
     
     
         5 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody or fragment thereof is monoclonal. 
     
     
         6 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody or fragment thereof is mammalian. 
     
     
         7 . The isolated antibody or fragment thereof of  claim 1 , wherein the antibody or fragment thereof is human. 
     
     
         8 . A hybridoma cell which produces the antibody or fragment thereof according to  claim 5 . 
     
     
         9 . The isolated antibody or fragment thereof of  claim 1 , wherein the fragment is selected from the group consisting of a Fab fragment; a F(ab′) 2 fragment; a Fv fragment; and a single chain fragment. 
     
     
         10 . The isolated antibody or fragment thereof of  claim 1 , further comprising a detectable substance coupled to the antibody. 
     
     
         11 . The isolated antibody or fragment thereof of  claim 10 , wherein the detectable substance is selected from the group consisting of an enzyme; a fluorescent label; a radioisotope; and chemiluminescent label. 
     
     
         12 . The isolated antibody or fragment thereof of  claim 10 , which specifically binds to neurogranin in an ELISA. 
     
     
         13 . The isolated antibody or fragment thereof of  claim 10 , which specifically binds to neurogranin in a competitive-binding assay. 
     
     
         14 . The isolated antibody or fragment thereof of  claim 10 , which specifically binds to neurogranin in a radioimmunoassay. 
     
     
         15 . The isolated antibody or fragment thereof of  claim 10 , which specifically binds to neurogranin in a fluorescence-activated cell sorting (FACS) assay. 
     
     
         16 . A kit for detecting neurogranin comprising: (a) the isolated antibody of  claim 1 ; and (b) at least one component to detect binding of the isolated antibody to neurogranin. 
     
     
         17 . An isolated antibody obtained from an animal that has been immunized with neurogranin, wherein the antibody specifically binds to an antigenic epitope-bearing polypeptide fragment of neurogranin. 
     
     
         18 . A monoclonal antibody designated 30.5.2 that specifically binds neurogranin. 
     
     
         19 . An anti-neurogranin monoclonal antibody which is produced by the hybridoma designated 30.5.2. 
     
     
         20 . A polynucleotide aptamer that specifically binds neurogranin. 
     
     
         21 . The polynucleotide aptamer of  claim 20 , wherein the neurogranin is human neurogranin. 
     
     
         22 . The polynucleotide aptamer of  claim 20 , wherein the aptamer binds to neurogranin with a Kd of less than about 1000 nM. 
     
     
         23 . The polynucleotide aptamer of  claim 20 , wherein the aptamer binds to neurogranin with a Kd of less than about 100 nM. 
     
     
         24 . The polynucleotide aptamer of  claim 20 , wherein the aptamer binds to neurogranin with a Kd of less than about 20 nM. 
     
     
         25 . The polynucleotide aptamer of  claim 20 , which consists of about 10 to about 100 nucleotides. 
     
     
         26 . The polynucleotide aptamer of  claim 25 , which consists of about 20 to about 80 nucleotides. 
     
     
         27 . The polynucleotide aptamer of  claim 26 , which consists of about 30 to about 50 nucleotides. 
     
     
         28 . The polynucleotide aptamer  claim 20 , which is an RNA aptamer. 
     
     
         29 . The polynucleotide aptamer of  claim 28 , comprising a nucleotide sequence at least 80% identical to any one of SEQ ID NOS: 4-6 or a fragment thereof of at least ten contiguous nucleotides. 
     
     
         30 . The polynucleotide aptamer of  claim 29 , comprising a nucleotide sequence at least 90% identical to any one of SEQ ID NOS: 4-6 or a fragment thereof of at least ten contiguous nucleotides. 
     
     
         31 . The polynucleotide aptamer of  claim 30 , comprising a nucleotide sequence at least 95% identical to any one of SEQ ID NOS: 4-6 or a fragment thereof of at least ten contiguous nucleotides. 
     
     
         32 . The polynucleotide aptamer of  claim 31 , comprising the nucleotide sequence of any one of SEQ ID NOS: 4-6 or a fragment thereof of at least ten contiguous nucleotides. 
     
     
         33 . The polynucleotide aptamer of  claim 31 , consisting of the nucleotide sequence of any one of SEQ ID NOS: 4-6 or a fragment thereof of at least ten contiguous nucleotides. 
     
     
         34 . The polynucleotide aptamer of  claim 20 , further comprising at least one modified internucleotide linker. 
     
     
         35 . The polynucleotide aptamer of  claim 20 , comprising at least one terminal blocker. 
     
     
         36 . The polynucleotide aptamer of  claim 20 , which is linked to a conjugate. 
     
     
         37 . A polynucleotide encoding the polynucleotide aptamer of  claim 20 . 
     
     
         38 . A vector comprising the polynucleotide of  claim 37 . 
     
     
         39 . A cell comprising the polynucleotide aptamer of  claim 20 . 
     
     
         40 . (canceled) 
     
     
         41 . A cell comprising the polynucleotide of  claim 37 . 
     
     
         42 . A cell comprising the vector of  claim 38 . 
     
     
         43 . A method of diagnosing a disease or disorder associated with neurogranin in a subject, comprising measuring the level of neurogranin in the subject by binding neurogranin with the polynucleotide aptamer of  claim 20  and determining the amount of aptamer bound to neurogranin. 
     
     
         44 . The method of  claim 43 , wherein the binding occurs in a sample obtained from the subject. 
     
     
         45 . A method for determining the amount of neurogranin in a sample comprising the step of detecting a peptide specific to neurogranin using a triple quadrupole mass spectrometer and multiple reaction monitoring, wherein the peptide specific to neurogranin comprises SEQ ID NO:7.