Anti-Nogo-A antibodies and methods of making and using thereof
Provided are novel human-derived monoclonal neutralizing Nogo-A specific antibodies as well as fragments, derivatives and variants thereof as well as methods related thereto. Polynucleotides, vectors, host cells and kits related to the Nogo-A specific antibodies are also provided. The antibodies, immunoglobulin chain(s), as well as binding fragments, derivatives and variants thereof can be used in pharmaceutical and diagnostic compositions for Nogo-A targeted immunotherapy and diagnosis, respectively.
1 . A recombinant monoclonal anti-Nogo-A antibody or an antigen-binding fragment thereof, comprising a variable heavy chain (VH) and a variable light chain (VL), wherein
(a) the VH comprises a VH complementary determining region 1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 3, a VH complementary determining region 2 (VH-CDR2) comprising the amino acid sequence of SEQ ID NO: 4, and a VH complementary determining region 3 (VH-CDR3) comprising the amino acid sequence of SEQ ID NO: 5; and
(b) the VL comprises a VL complementary determining region 1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 8, a VL complementary determining region 2 (VL-CDR2) comprising the amino acid sequence of SEQ ID NO: 9, and a VL complementary determining region 3 (VL-CDR3) comprising the amino acid sequence of SEQ ID NO: 10.
2 . The antibody or antigen-binding fragment thereof of claim 1 , wherein
the VH comprises the amino acid sequence of SEQ ID NO: 2, and
the VL comprises the amino acid sequence of SEQ ID NO: 7.
3 . The antibody or antigen-binding fragment thereof of claim 1 , further comprising an immunoglobulin heavy chain constant region and an immunoglobulin light chain constant region.
4 . The antibody or antigen-binding fragment thereof of claim 3 , comprising a mutated Fc region with reduced effector functions as compared to a wildtype IgG Fc region.
5 . The antibody or antigen-binding fragment thereof of claim 3 , wherein the immunoglobulin heavy chain constant region is of IgG type.
6 . The antibody or antigen-binding fragment thereof of claim 5 , wherein the immunoglobulin heavy chain constant region is of IgG4 type.
7 . The antibody or antigen-binding fragment thereof of claim 6 , wherein the immunoglobulin heavy chain constant region comprises an S228P mutation, numbered according to the Kabat numbering system.
8 . The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody is a human-derived antibody.
9 . The antibody or antigen-binding fragment thereof of claim 1 , wherein the antigen-binding fragment thereof is selected from the group consisting of a single chain Fv fragment (scFv), an F(ab′) fragment, an F(ab) fragment, an F(ab′) 2 fragment, an Fd, an Fv, a single-chain antibody, and a disulfide-linked Fv (sdFv); and/or the antibody is a chimeric murine-human antibody.
10 . The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof
(i) is detectably labeled with a label selected from the group consisting of an enzyme, a radioisotope, a fluorescent compound, a chemiluminescent compound, a bioluminescent compound, a tag, a flag, or a heavy metal;
(ii) is conjugated to polyethylene glycol.
11 . The anti-Nogo-A antibody or an antigen-binding fragment thereof of claim 1 , wherein the anti-Nogo-A antibody or antigen-binding fragment is produced by a method comprising (a) culturing an isolated host cell comprising one or more polynucleotide(s) encoding the antibody or antigen-binding fragment thereof of claim 1 in a culture medium; and (b) isolating the antibody or antigen-binding fragment thereof expressed by the host cell from the culture medium.
12 . The antibody or antigen-binding fragment thereof of claim 11 , wherein the antibody or antigen-binding fragment thereof
(i) is detectably labeled with a label selected from the group consisting of an enzyme, a radioisotope, a fluorescent compound, a chemiluminescent compound, a bioluminescent compound, a tag, a flag, or a heavy metal;
or
(ii) is conjugated to polyethylene glycol.
13 . A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.
14 . A diagnostic kit comprising the antibody or antigen-binding fragment thereof of claim 1 .
15 . A method for promoting neurite outgrowth, vascular repair and/or motor functional recovery in an injury of the peripheral nervous system (PNS) and/or central nervous system (CNS) in a subject in need thereof, comprising administering the antibody or antigen-binding fragment thereof of claim 1 to the subject.
16 . The method of claim 15 , wherein the injury of the PNS and/or CNS is cranial trauma, cerebral trauma, spinal trauma, stroke, traumatic brain injury, or a demyelinating disease.
17 . The method of claim 15 , wherein the injury is spinal cord injury.
18 . A method for in vivo detection of Nogo-A in a subject, comprising administering the detectably labeled antibody or antigen-binding fragment thereof of claim 10 to the subject; and detecting presence of an immunocomplex of the detectably labeled antibody or antigen-binding fragment thereof and Nogo-A in vivo by immunofluorescent imaging.