NEUTRALIZING MONOCLONAL ANTIBODIES AGAINST THE NOGO-66 RECEPTOR (NgR) AND USES THEREOF

Abstract

The subject invention relates to isolated proteins, particularly monoclonal antibodies, which bind to the Nogo-66 receptor. Specifically, these antibodies have the ability to inhibit the binding of the natural ligand of the Nogo-66 receptor and neutralize the Nogo-66 receptor. These antibodies or portions thereof of the invention are useful for detecting NgR and for inhibiting NgR activity, for example in a human suffering from a disorder in which NgR or Nogo-66 activity is detrimental.

1 . An isolated binding protein that specifically interacts to at least one epitope of a Nogo-66 receptor. 
     
     
         2 . The isolated binding protein of  claim 1 , wherein the isolated protein is a monoclonal neutralizing antibody or antigen binding fragment thereof. 
     
     
         3 . The binding protein according to  claim 2  wherein said antigen binding fragment comprises a VH and a VL domain. 
     
     
         4 . The neutralizing antibody according to  claim 2  wherein said neutralizing antibody diminishes the ability of Nogo-66 to bind to its receptor. 
     
     
         5 . The neutralizing antibody according to  claim 2  wherein said neutralizing antibody is capable of neutralizing Nogo-66. 
     
     
         6 . The antibody according to  claim 2  wherein the Nogo66 receptor is selected from human, cynomolgus monkeys, rat, chick, frog, and fish. 
     
     
         7 . The antibody according to  claim 2  wherein the Nogo66 receptor polypeptide shares 90% homology to the amino acid sequence selected from SEQ ID NO:1a and SEQ ID NO:2a. 
     
     
         8 . The antibody according to  claim 2  wherein the Nogo66 receptor is encoded by a nucleic acid that shares 90% homology to the nucleic acid sequence SEQ ID NO:1b and SEQ ID NO:2b. 
     
     
         9 . The antibody of  claim 3  that has at least 90% amino acid sequence identity with a sequence comprising a heavy chain variable region (VH region) comprising the sequence of SEQ ID NO:3. 
     
     
         10 . The antibody of  claim 3  that has at least 90% amino acid sequence identity with a sequence comprising a light chain variable region (VL region) comprising the sequence of SEQ ID NO:4. 
     
     
         11 . The antibody of  claim 3  that has at least 90% amino acid sequence identity with a sequence comprising a heavy chain variable region (VH region) comprising the sequence of SEQ ID NO:5. 
     
     
         12 . The antibody of  claim 3  that has at least 90% amino acid sequence identity with a sequence comprising a light chain variable region (VL region) comprising the sequence of SEQ ID NO:6. 
     
     
         13 . The antibody according to  claim 3  that has a dissociation constant (K d ) ranges from 10 −7  M to 10 −13  M. 
     
     
         14 . The antibody according to  claim 3  has an on rate (K on ) ranging from 10 3  M −1 s −1  to 10 7 M-1s −1 . 
     
     
         15 . The antibody according to  claim 3  that has an off rate (K off ) ranging from 10 −3  s −1  to at least about 10 −6 s −1 . 
     
     
         16 . The antibody according to  claim 3  that has an IC 50  value ranging from 10 −6  s −1  to at least about 10 −12 s −1 . 
     
     
         17 . The antibody according to  claim 3  that binds to human NgR with an EC 50  ranging from 1×10 −9  M, to 1×10 −11  M. 
     
     
         18 . The antibody according to  claim 3  that binds to human NgR wherein the antibody is glycosylated. 
     
     
         19 . The antibody or antigen-binding fragment of  claim 3 , wherein said antibody or antigen-binding fragment is a mouse antibody, a humanized antibody, a fully human, a chimeric antibody, an antigen-binding fragment of a humanized antibody, or an antigen-binding fragment of a chimeric antibody. 
     
     
         20 . The antibody or antigen-binding fragment of  claim 3 , wherein said antibody or antigen-binding fragment is an antigen-binding fragment selected from the group consisting of a Fab fragment, a Fab′ fragment, a F(ab′) 2  fragment and a Fv fragment. 
     
     
         21 . A monoclonal antibody that specifically binds to at least one epitope of a Nogo-66 receptor, wherein said monoclonal antibody is the monoclonal antibody secreted by hybridoma cell line ATCC NO. PTA-8383 and ATCC NO. PTA-8384. 
     
     
         22 . (canceled) 
     
     
         23 . The monoclonal antibody of  claim 21 , wherein the binding results in inactivation of the Nogo-66 receptor. 
     
     
         24 . A hybridoma cell line that produces a monoclonal antibody, which specifically binds to at least one epitope of a Nogo-66 receptor. 
     
     
         25 . The hybridoma cell line of  claim 24 , wherein the hybridoma is selected from the group consisting of mouse, rat, sheep, pig, cattle, goat, horse, and human hybridoma. 
     
     
         26 . (canceled) 
     
     
         27 . The monoclonal antibody of  claim 22 , wherein the binding results in inactivation of the Nogo-66 receptor. 
     
     
         28 . (canceled) 
     
     
         29 . The hybridoma cell line of  claim 27 , wherein the hybridoma is selected from the group consisting of mouse, rat, sheep, pig, cattle, goat, horse, and human hybridoma. 
     
     
         30 . (canceled) 
     
     
         31 . A monoclonal neutralizing antibody or antigen-binding fragment thereof, having at least one characteristic selected from the group consisting of:
 a) binding to mammalian NgR with affinity in the nM range or less;   b) functionally antagonizing in vitro Nogo-66 binding in neurite outgrowth assay with Nm or less efficacy;   c) functionally antagonizing in vitro Nogo-66 binding in neurite outgrowth assay with nM or less efficacy;   d) in vivo inducing sprouting in the optic nerve crush model; or   e) relieving in vivo experimental spinal cord injury.   
     
     
         32 . An isolated nucleic acid encoding the monoclonal neutralizing antibody or antigen-binding fragment according to  claim 3 . 
     
     
         33 . A vector comprising the isolated nucleic acid of  claim 28 , wherein said vector is selected form the group consisting of pcDNA; pTT; pTT3; pEFBOS; pBV; pJV; and pBJ. 
     
     
         34 . A host cell transformed with the vector according to  claim 33 , wherein the host cell is selected from the group consisting of protist cell, animal cell, plant cell and fungal cell. 
     
     
         35 . The host cell of  claim 30  wherein the animal cell is a mammalian cell selected from the group comprising HEK293, CHO and COS. 
     
     
         36 . A host cell transformed with the vector according to  claim 33 , wherein the host cell is a eukaryotic cell. 
     
     
         37 . A method of producing the binding protein according to  claim 1  that binds human and/or rat NgR, comprising culturing a host cell in a culture medium under conditions sufficient to produce a binding protein that binds human and/or rat NgR. 
     
     
         38 . A pharmaceutical composition comprising the monoclonal antibody or antigen-binding portion of  claim 3  and a pharmaceutically acceptable carrier. 
     
     
         39 . A method for decreasing Nogo-66 binding to Nogo-66 receptor in a subject in need thereof, comprising the step of administering to the subject an isolated protein of  claim 3 . 
     
     
         40 . A method of reversing Nogo-66-induced neurite outgrowth inhibition comprising the step of using the isolated protein of  claim 3 . 
     
     
         41 . A method of treating a subject for a disorder associated with NgR activity comprising the step of administering alone or in combination with other therapeutic agents the antibodies of  claim 3 . 
     
     
         42 . The method of  claim 41  wherein the disorder comprises neurological diseases selected from the group comprising Amytropic Lateral Sclerosis; Brachial Plexus Injury, Brain Injury, including traumatic brain injury, Cerebral Palsy, Friedrich's Ataxia, Guillain Barre, Leukodystrophies, Multiple Sclerosis, Post Polio, Spina Bifida, Spinal Cord Injury, Spinal Muscle Atrophy, Spinal Tumors, Stroke, and Transverse Myelitits, dementia, senile dementia, mild cognitive impairment, Alzheimer-related dementia, Huntington's chorea, tardive dyskinesia, hyperkineasias, manias, Morbus Parkinson, steel-Richard syndrome, Down's syndromes, myasthenia gravis, nerve trauma, vascular amyloidosis, cerebral hemorrhage with amyloidosis, brain inflammation, Friedrich's ataxia, acute confusion disorder, amyotrophic lateral sclerosis, glaucoma, and Alzheimer's disease. 
     
     
         43 . (canceled) 
     
     
         44 . A method for reducing human NgR activity in a subject suffering from a disorder in which NgR activity is detrimental, comprising administering to the subject the antibodies of  claim 3 , alone or in combination with other therapeutic agents, such that human NgR activity in the subject is reduced.