CN114409779A

CN114409779A - NogoA antibody and application thereof in spinal cord injury

Info

Publication number: CN114409779A
Application number: CN202210101569.1A
Authority: CN
Inventors: 王秀娟
Original assignee: Tianjin Changhe Biological Technology Co ltd
Current assignee: Tianjin Changhe Biological Technology Co ltd
Priority date: 2022-01-27
Filing date: 2022-01-27
Publication date: 2022-04-29
Anticipated expiration: 2042-01-27
Also published as: CN114409779B

Abstract

The invention relates to the field of biomedicine, in particular to a NogoA antibody and application thereof in spinal cord injury. The NogoA antibody prepared by the invention has high affinity and good specificity, and does not have cross reaction with NogoB and NogoC. In particular, an important advantage of the antibody is that it has the activity and physical stability of binding and neutralizing NogoA, and thus is preferably used as an antibody drug, which has excellent application prospects in the treatment of spinal cord injury.

Description

NogoA antibody and application thereof in spinal cord injury

Technical Field

The invention relates to the field of biomedicine, in particular to a NogoA antibody and application thereof in spinal cord injury.

Background

The term "Spinal Cord Injury" (Spinal Cord Injury) refers to Spinal Cord damage caused by trauma (e.g., traffic collision), disease, or degeneration (e.g., cancer). There is currently no reliable estimate of global prevalence, but mostly due to trauma, although the proportion of non-traumatic spinal cord injuries presents an increasing trend. The symptoms of spinal cord injury depend on the severity of the injury and the location of the damaged spinal cord. Symptoms may include partial or complete loss of sensory function or motor control of the arms, legs, and/or body. The most severe spinal cord injuries affect the body system that regulates urinary and fecal control, respiration, heart rate, and blood pressure. Most spinal cord injuries experience chronic pain and are at risk of developing secondary diseases, such as deep vein thrombosis, urinary tract infections, muscle spasms, osteoporosis, bed sores, chronic pain and respiratory complications, which can be debilitating or even life threatening. Emergency care, rehabilitation services and ongoing health maintenance are critical to the prevention and management of these diseases. The consequences are lifelong and devastating, which not only causes great pain to the patients, but also brings heavy burden to the family and the society.

It is presently believed that the major adverse factors affecting regeneration after spinal cord injury are myelin-associated inhibitory molecules and glial scars. NogoA, also known as Reticulon 4, is a protein encoded by the RTN4 gene in humans and has been identified as an inhibitor of the central nervous system-specific production of neurogenic muscle. During neurodevelopment, Nogo is predominantly expressed by neurons and provides inhibitory signals for migration and sprouting of CNS endothelial (apical) cells, thereby limiting vascular density. Studies have shown that blocking NogoA (from diseases such as multiple sclerosis) during neuronal injury will help protect or restore damaged neurons. Studies with this protein have shown great potential for treatment of autoimmune-mediated demyelination and regeneration of spinal cord injury.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

The invention relates to an NogoA antibody or an antigen-binding fragment thereof, which comprises a heavy chain complementary determining region CDR-VH1, CDR-VH2 and CDR-VH3 of which the amino acid sequences are shown as SEQ ID NO. 1-3 in sequence, and a light chain complementary determining region CDR-VL1, CDR-VL2 and CDR-VL3 of which the amino acid sequences are shown as SEQ ID NO. 4-6 in sequence.

In the present invention, the term "antibody" is a protein that binds to a specific antigen, and broadly refers to all proteins and protein fragments that comprise CDR regions, particularly full-length antibodies. The term "full-length antibody" includes both polyclonal and monoclonal antibodies, and the term "antigen-binding fragment" is a substance that comprises part or all of an antibody CDR, which lacks at least some of the amino acids present in the full-length chain but is still capable of specifically binding to an antigen. Such fragments are biologically active in that they bind to a target antigen and can compete with other antigen binding molecules (including whole antibodies) for binding to a given epitope. In some embodiments, the functional fragment of an antibody has the effect of specifically recognizing and binding NogoA. In one aspect, such fragments will comprise a single heavy chain and a single light chain, or portions thereof. Such fragments may be produced by recombinant nucleic acid techniques, or may be produced by enzymatic or chemical cleavage of antigen binding molecules, including intact antibodies.

The term "complementarity determining regions" or "CDRs" refers to the highly variable regions of the heavy and light chains of immunoglobulins, as defined by Kabat et al (Kabat et al, Sequences of proteins of immunological interest,5th Ed "US Department of health and Human Services, NIH,1991, and later versions) in the present invention. There are three heavy chain CDRs (HCDRs) and three light chain CDRs (LCDRs). Herein, the terms "CDR" and "CDRs" are used to refer to a region comprising one or more, or even all, of the major amino acid residues that contribute to the binding affinity of an antibody to the antigen or epitope it recognizes, depending on the circumstances.

Alternatively, the NogoA antibody or antigen-binding fragment thereof as described above, comprising a heavy chain variable region HCVR as shown in SEQ ID NO. 7 and a light chain variable region LCVR as shown in SEQ ID NO. 8.

Also within the scope of the invention are variants of antibodies or antigen-binding fragments thereof comprising heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3, and light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, and HCVR and LCVR, wherein the sequences of said HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, and variants of HCVR and LCVR, respectively, comprise a mutation of up to 3 amino acids (e.g., a substitution, deletion or addition of 1, 2 or 3 amino acids, or any combination thereof) as compared to any of the sequences set forth in SEQ ID NOs 1-8; preferably sequences having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity.

The term "antigen-binding fragment" includes antigen-compound-binding fragments of these antibodies, including Fab, F (ab ')2, Fd, Fv, scFv, minimum recognition units of antibodies, and single chain derivatives of these antibodies and fragments, such as scFv-Fc and the like, preferably F (ab')₂、Fab、scFv。

Preferably, the NogoA antibody or antigen binding fragment thereof as described above, said antibody having a constant region.

Further preferably, the heavy chain constant region sequence is selected from the group consisting of constant region sequences of any one of IgG, IgA, IgM, IgE, IgD; wherein the IgG may be further divided into subclasses, e.g., selected from IgG1, IgG2, IgG3, IgG 4.

Further preferably, the light chain constant region is a kappa or lambda chain;

further preferably, the species of the constant region is from a source selected from the group consisting of cow, horse, pig, sheep, mouse, dog, cat, rabbit, camel, donkey, deer, mink, chicken, duck, goose, and human.

Alternatively, the antibody or antigen-binding fragment thereof as described above is a mouse antibody, a human-mouse chimeric antibody, or a humanized antibody.

According to a further aspect of the invention, it also relates to an isolated nucleic acid encoding a NogoA antibody or antigen binding fragment thereof according to the above.

The invention also relates to a vector comprising a nucleic acid as described above. The term "vector" refers to a nucleic acid delivery vehicle into which a polynucleotide can be inserted. When a vector is capable of expressing a protein encoded by an inserted polynucleotide, the vector is referred to as an expression vector. The vector may be introduced into a host cell by transformation, transduction, or transfection, and the genetic material elements carried thereby are expressed in the host cell. Vectors are well known to those skilled in the art and include, but are not limited to: a plasmid; phagemid; a cosmid; artificial chromosomes such as Yeast Artificial Chromosomes (YACs), Bacterial Artificial Chromosomes (BACs), or artificial chromosomes (PACs) derived from P1; bacteriophage such as lambda phage or M13 phage, animal virus, etc. Animal viruses that may be used as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpes viruses (e.g., herpes simplex virus), poxviruses, baculoviruses, papilloma viruses, papilloma polyoma vacuolatum viruses (e.g., SV 40). In some embodiments, regulatory elements commonly used in genetic engineering, such as enhancers, promoters, Internal Ribosome Entry Sites (IRES), and other expression control elements (e.g., transcription termination signals, or polyadenylation signals and poly-U sequences, etc.) are included in the vectors of the present invention.

The invention also relates to a host cell comprising a nucleic acid as described above or a vector as described above. The term "host cell" refers to a cell which can be used for introducing a vector, and includes, but is not limited to, prokaryotic cells such as Escherichia coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus, insect cells such as S2 Drosophila cells or Sf9, or animal cells such as fibroblast, CHO cells, COS cells, NSO cells, HeLa cells, BHK cells, HEK 293 cells or human cells. The host cell is preferably a eukaryotic cell, more preferably a mammalian cell.

The invention also relates to a pharmaceutical composition comprising a NogoA antibody, or antigen-binding fragment thereof, as described above, and one or more of a pharmaceutically acceptable excipient, diluent or carrier.

The term "pharmaceutically acceptable excipient, diluent or carrier" refers to an excipient, diluent or carrier that is pharmacologically and/or physiologically compatible with the subject and active ingredient, which is well known in the art, including but not limited to: pH regulator, surfactant, adjuvant, and ionic strength enhancer. For example, pH adjusting agents include, but are not limited to, phosphate buffers; surfactants include, but are not limited to, cationic, anionic or nonionic surfactants, such as Tween-80; ionic strength enhancers include, but are not limited to, sodium chloride.

The invention also relates to the use of an NogoA antibody or antigen-binding fragment thereof as described above for the preparation of a medicament for the treatment of spinal cord injury.

Has the advantages that:

the NogoA antibody prepared by the invention has high affinity and good specificity, and does not have cross reaction with NogoB and NogoC. In particular, an important advantage of the antibody is that it has the activity and physical stability of binding and neutralizing NogoA, and thus is preferably used as an antibody drug, which has excellent application prospects in the treatment of spinal cord injury.

Detailed Description

Reference will now be made in detail to embodiments of the invention, one or more examples of which are described below. Each example is provided by way of explanation, not limitation, of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. For instance, features illustrated or described as part of one embodiment, can be used on another embodiment to yield a still further embodiment.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Embodiments of the present invention will be described in detail with reference to examples.

EXAMPLE 1 preparation of antibodies

1. Protein design and expression

The Nogo gene has three isomers, namely nogoA, nogoB and nogoC according to the difference of a promoter and a shearing mode, primers are designed on two sides of a 66 amino acid reading frame according to a known sequence of a cDNA Genbank (Accession No: AJ242963) of Nogo, an upstream primer AGCTTTAGCATATATAAG GC and a downstream primer TTAGGAATCAACTAAATCATCAAC are used for amplifying and fishing related genes from rat brain tissues, a Flag-his tag is fused on the basis of a NogoA (aa1026-1091, the sequence of which is peculiar to the nogoA) protein, the obtained product is cloned on a pTT5 vector (Biovector, Cat #:102762), and the product is subjected to transient expression in 293 cells or stable expression and purification in CHO-S to obtain the antigen and the protein for detection.

2. Protein purification

The cell expression fluid is centrifuged at high speed, the supernatant is collected, and the precipitate is discarded. HisTrap FF pre-packed columns were equilibrated with Phosphate Buffered Saline (PBS) at 5-10 column volumes. The cell expression supernatant was loaded at a rate of 2 ml/min. Washing the pre-packed column with PBS until mAu reading reaches a base line, then eluting the target protein with 20mM, 50mM and 250mM of imidazole in sequence and collecting, finally transferring the target protein eluted by 300mM of imidazole into a concentration tube, centrifuging, changing the solution, and replacing the target protein into a PBS solution for storage for subsequent experiments.

3. Antigen immunization and monoclonal preparation

Injecting conjugate of human NogoA and mouse antibody Fc Fragment (Fragment crystalline) intraperitoneally into BALB/c mouse, 100 μ g/200 μ l/mouse once per week, taking mouse tail blood every week after 3 weeks of immunization and detecting expression of NogoA antibody in serum; mice with high expression of NogoA antibody in serum (dotblot assay) were selected for splenocytes and fused with SP2/0 myeloma cells to form fusions.

The cell fusion method comprises the following steps:

cell fusion was carried out using 50% PEG of molecular weight 1450 as a fusogenic agent, and was carried out according to the conventional method, which was carried out as follows:

take 1X 10⁷The SP2/0 myeloma cells were mixed with the spleen cells, resuspended in the basic medium, and washed three times at 2000r/min and 5 min. The sterilized absorbent paper was removed from the clean bench, and the supernatant of a 50mL centrifuge tube containing the mixture of myeloma cells and spleen cells was poured off, and the tube was inverted on the absorbent paper to remove water droplets. Using a small aluminum pot to contain 2/3 volumes of water, heating the water on an electric furnace to 37 ℃, and then putting the water into a superclean bench; from CO₂Taking out 50% PEG incubated to 37 ℃ from the incubator, sucking 0.8mL by using a 1mL suction tube, holding a 50mL centrifuge tube filled with mixed cells by hand, placing the centrifuge tube in a small aluminum pot with water bath at 37 ℃, slowly adding PEG to the mixed cells while gently stirring, after continuing for 2min, inserting the centrifuge tube into a centrifuge tube rack, removing the small aluminum pot, and removing CO from the centrifuge tube₂The incubator takes out 50mL of basic culture solution incubated to 37 ℃, sucks 10mL of basic culture solution by a pipette, slowly adds the basic culture solution to the fused cells while gently stirring to disperse cell masses, firstly adds 1mL of basic culture solution, then adds 2mL of basic culture solution, then adds 3mL of basic culture solution, finally adds the rest 4mL of basic culture solution, and after the first 10mL of basic culture solution is added, then adds the rest 40mL of basic culture solution along the tube wall, after the addition is finished, screws down the cover, and repeatedly turns over for several times to uniformly mix the cells. 1500r/min, centrifuge for 5min, discard the supernatant, resuspend the fused cells in 72mL of complete medium with resuspended feeder cells. Note: the movement needs to be light, the cells will be gently stirred up by the pipette without blowing. The resuspended cells were added dropwise to a 96-well cell culture plate at 2 drops/well and CO was added₂Culturing in an incubator. Microcolonies were observed on day 4 post-fusion; supplementing 1 drop/hole of complete culture solution on day 7; on about day 10, colonies grew to 1/4% and the culture medium turned yellow, and antibody detection was performed.

Screening and cloning of hybridoma positive clones:

and (4) observing the culture plate under an inverted microscope after 10-14 days of fusion, and marking a small point above the culture plate cover with the colony-appearing hole by using a marker pen to mark the position of the colony so as to facilitate sampling and number matching of the original hole during detection.

Selecting a fusion expressing the NogoA antibody in the culture supernatant for monoclonal; and (3) selecting a monoclonal hybridoma cell strain expressing the NogoA antibody for expanding culture, collecting cell culture solution after culturing for 7-10 days, and purifying to obtain the NogoA antibody.

Example 2 antibody assay

1) Determination of antibody Activity

1. The probe with the surface coupled with the protein ProteinA is soaked in 250 μ L of buffer K (PBS + 0.002% Tween 20+ 0.02% BSA) for 10 minutes;

2. preparing the antibody into a working solution of 5 mu g/mL by using buffer K;

3. preparing the recombinant human NogoA protein into working solutions with four concentrations of 10 mug/mL, 5 mug/mL, 2.5 mug/mL and 0 mug/mL respectively by using buffer K;

4. according to the indication of a Gator non-labeled analyzer (star child medical technology, CAT #: Gator), reagents are added, the affinity data is shown in table 1, and the 13 cloned antibodies with good primary screening effect are detected at this time. The results of the assay showed that the binding kinetic constants of most anti-NogoA antibodies to NogoA recombinant proteins were on the pM scale.

TABLE 1

Clone editingNumber (C)	K_D(M)，×10E^-10(M)
		NogoAB2-3	1.43
NogoAB2-4	0.057
		NogoAB2-6	3.72
NogoAC3-3	5.01
		NogoAC4-6	2.11
NogoAD1-1	0.082
		NogoAD5-3	0.295
NogoAF3-2	3.22
		NogoAG3-1	4.19
NogoAG6-4	7.28
		NogoAH2-2	0.284
NogoAH4-1	7.98
		NogoAH2-2	19.29

2) Physical stability testing of antibodies

Four antibodies, NogoAB2-4, NogoAD1-1, NogoA D5-3 and NogoAH2-2, with better effect were selected, and the thermal stability of the antibodies was examined by DSC (Differential scanning calorimetry). Differential thermal scanning (DSC) measures the thermodynamic changes of molecular interactions by inducing changes in macromolecules by increasing or decreasing temperature, and is the first technique to characterize molecular thermal stability.

The four antibodies were diluted to the same concentration and the detection was performed according to MicroCal VP-Capillary DSC (Malvern) instructions. Before detection, each sample and blank buffer solution are degassed for 1-2 min by a vacuum degasser. Add 500. mu.l sample or blank buffer per well of the sample plate (instrument load 300. mu.l). Finally, 14% Decon 90 and ddH were added to the two pairs of plates₂And O, for cleaning, and sleeving a plastic soft cover plate after the sample plate is loaded. The scanning temperature starts from 25 ℃ and ends at 100 ℃ and the scanning speed is 60 ℃/h. Specific results are shown in table 2.

TABLE 2

Antibodies are relatively stable in proteins, and their Tm values are substantially around 70 degrees. However, the better the thermal stability of the antibody protein used as a drug, the less likely it is to be degraded during transportation/storage. From the experimental results, NogoAD1-1 showed better thermostability compared to other antibodies.

Monitoring the purity of the sample by SEC-HPLC, inspecting the periodic stability under a certain concentration condition, controlling the concentration of the sample to be about 1mg/ml, and detecting the stability of the antibody in a PBS system by repeated freeze thawing for 3 times (the freezing temperature is-70 ℃ every time, and the antibody is naturally thawed at room temperature). Antibody purity was checked using an Xbridge protein BEH SEC200A (Waters) HPLC column. As shown in Table 3, in PBS, the antibody NogoAD1-1 showed better stability than the other antibodies.

TABLE 3

NogoAB2-4(△％)	NogoAD1-1(△％)	NogoAD5-3(△％)	NogoAH2-2(△％)
				2.9	1.2	5.4	5.2

Remarking: delta% indicates the rate of change of HPLC purity reduction

The NogoA D1-1 antibody performed best overall after comprehensive judgment, and the amino acid sequence VH of the sequenced heavy chain variable region is shown as SEQ ID NO. 7, and the amino acid sequence VL of the light chain variable region is shown as the sequence SEQ ID NO. 8. The antibody type is IgG.

Heavy Chain variable region:(SEQ ID NO:7)

EVLLVESQQTPVKPGGSLKASCAGSQSVFLTAAYSWVRPTPEKLLEWVATISYGSPNTYYPDSVKGRYNEKFKGKATLTVETSSSTAYMQLSSLTSEDTAVYFCARRSYGEDSWFSYWGQGTLVTVSS

Light Chainvariable region:(SEQ ID NO:8)

DIVMTQSPPSLAISVGQKVTMSCKSSQSILNSSAQRFCLAWYSQKPGESPKDLVYVYSSRDSGVPDRFIGSGTAFTFTLTISSLQVEDLADYFCQNHFSIPAMFGAGTKLESS

Example 3 Tail intravenous injection of antibodies accelerates recovery from spinal cord injury

The experimental method is as follows:

1. laboratory animal

The experimental animals are SD male rats of 250-280 g, purchased from Beijing Wittiulihua laboratory animals technology Co. The animals were kept in a constant temperature environment of 23 deg.C with humidity of 40% and circadian rhythm of 12/12 hr. The mouse food and water are sufficiently supplied.

2. And (5) establishing a rat spinal cord impact injury model.

See the method of Wamil et al [ Wamil et al Proc NatlAcad Sci USA, 1998; 95(22): 13188-. The device consists of a striking rod, a peripheral sleeve, a hammer, an external fixing frame and the like. The diameter of the head end of the striker is 2.3mm, the height is 10mm, the weight of the hammer is 10g, the falling height is 1.25cm, and the impact force of the falling body caused by injury is 10 multiplied by 1.25g cm. Animals were weighed at 23 ℃ and anesthetized by intraperitoneal injection with 5% chloral hydrate (0.75ml/100g) and fixed in a rat holder. Incising the skin of the back and the subcutaneous tissue of the spinal cord along the median line for about 4cm, positioning the spinous process of the T11 vertebral body, separating the muscles at two sides of the T11, 10 and 9 vertebral bodies bluntly, and opening and fixing the opening device to expose the three vertebral bodies for laminotomy. The head end of the striker is arranged on a T10 section of spinal cord, the outer sleeve tube is fixedly struck, the striking hammer freely falls along the outer sleeve tube from the height of 1.25cm to impact the striker, the T10 section of spinal cord contusion is caused, and the forward flapping, the tail flicking and the apnea of the body are seen in the operation to indicate the success of model building. Stopping bleeding with gelatin sponge, suturing muscle, subcutaneous tissue and skin layer by layer, feeding water after operation, and raising in cages. The bladder squeezes urine 4-5 times a day, and the urine lasts for 1-2 w until the micturition reflex is recovered.

3. Experimental groups and antibody administration strategies

The method comprises the steps of selecting a tail vein injection antibody after a spinal cord injury model is successfully established for 24h, 3d and 7d, and injecting the antibody three times in total. The experiment was divided into groups of 8 animals per group.

Control: a blank control group, PBS + IgG is injected, and the antibody concentration is 10 mug/g;

group1, tail vein injection of PBS + NogoAD1-1 antibody at an antibody concentration of 5 μ g/g (based on rat body weight);

group2, tail vein injection of PBS + NogoAD1-1 antibody, 10. mu.g/g antibody concentration;

group3, tail vein injection of PBS + NogoAD1-1 antibody, antibody concentration 15 μ g/g;

4. assessment of efficacy

4.1BBB scoring was performed by double blind experiments, and hind limb functions including joint movement, walking ability, coordination and limb stability were evaluated separately for experimental rats. BBB scores were performed in groups at 30d post spinal cord injury.

Statistical analysis data processing using SPSS15.0 software, results in

And (4) showing. BBB scores and cell counts were determined using paired t-tests or one-way analysis of variance (ANOVA). p is a radical of<0.05 is statistically significant.

Rat hind limb BBB scores are referenced in table 4.

TABLE 4 BBB sports function scoring table

Scores of each preoperative group were 21 points, and scores of each group were 0 point after injury. After 30 days of rest, the Control group is recovered initially, and the scores of the rest groups are obviously improved relative to the score of the Control group.

TABLE 5

Note: p < 0.05, vs Control.

From the results, the NogoA antibody prepared by the invention can effectively inhibit the activity of NogoA protein under the antibody concentration of 5 mu g/g or more, thereby accelerating the recovery of spinal cord injury and having good application prospect.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

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Claims

The NogoA antibody or an antigen-binding fragment thereof is characterized by comprising a heavy chain complementarity determining region CDR-VH1, CDR-VH2 and CDR-VH3 of which the amino acid sequences are shown as SEQ ID NO. 1-3 in sequence, and a light chain complementarity determining region CDR-VL1, CDR-VL2 and CDR-VL3 of which the amino acid sequences are shown as SEQ ID NO. 4-6 in sequence.
2. An NogoA antibody or antigen-binding fragment thereof according to claim 1, which comprises a heavy chain variable region HCVR as shown in SEQ ID No. 7 and a light chain variable region LCVR as shown in SEQ ID No. 8.
3. The NogoA antibody, or an antigen-binding fragment thereof, according to claim 1 or 2, wherein the antigen-binding fragment is F (ab')₂Fab, scFv and diabody.
4. An NogoA antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody has a constant region and the heavy chain constant region sequence is selected from the group consisting of the constant region sequences of any one of IgG, IgA, IgM, IgE, IgD; the light chain constant region is a kappa or lambda chain;

preferably, the species source of the constant region is selected from cattle, horses, pigs, sheep, mice, dogs, cats, rabbits, camels, donkeys, deer, mink, chickens, ducks, geese or humans.
5. The antibody or antigen-binding fragment thereof according to claim 4, which is a mouse antibody, a human-mouse chimeric antibody or a humanized antibody.
6. An isolated nucleic acid encoding the NogoA antibody or antigen binding fragment thereof according to any one of claims 1 to 5.
7. A vector comprising the nucleic acid of claim 6.
8. A host cell comprising the nucleic acid of claim 6 or the vector of claim 7.
9. A pharmaceutical composition comprising the NogoA antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, and one or more of a pharmaceutically acceptable excipient, diluent or carrier.
10. Use of an NogoA antibody or antigen binding fragment thereof according to any one of claims 1 to 5 in the manufacture of a medicament for the treatment of spinal cord injury.