Antibody specifically binding to alpha-synuclein and related product and application thereof

Abstract

The invention discloses an antibody specifically binding to alpha-synuclein and a related product and application thereof, and provides a brand new monoclonal antibody targeting to alpha-synuclein for the field, the monoclonal antibody can specifically bind to alpha-synuclein, has good specificity and affinity, and can be used for detecting alpha-synuclein. A foundation is laid for development of alpha-synuclein related detection and / or auxiliary detection products in the field, meanwhile, more diagnostic tools and treatment strategies are expected to be provided for diagnosis and / or treatment of alpha-synucleinopathies, and good application prospects are achieved.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to an antibody specifically binding to alpha-synuclein and related products and uses thereof. Background Art

[0002] Parkinson's disease (PD) is the second largest neurodegenerative disease. Its main pathological features are degeneration and loss of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies in the remaining neurons. In addition to motor symptoms such as muscle rigidity, bradykinesia, resting tremor, abnormal posture and gait, PD also presents non-motor disorders such as olfactory disorders, constipation, and rapid eye movement sleep disorders. Non-motor symptoms appear 10-20 years earlier than motor symptoms. When patients develop motor symptoms, their disease has reached an irreversible stage. Therefore, early diagnosis and treatment of PD are particularly important.

[0003] α-Synuclein (SNCA) is the main structural component of Lewy bodies, which are one of the pathological characteristics of PD and are formed by the aggregation of α-synuclein. In the brain tissue of PD patients, α-synuclein is abnormally expressed and aggregated, leading to biochemical reactions such as oxidative stress and the formation of oligomeric intermediate conformations, which play an important role in the pathogenesis of PD. α-Synuclein is a soluble protein expressed presynaptically and perinuclearly in the central nervous system. It is composed of 140 amino acids and has a molecular weight of 14 kDa. α-Synuclein is the most important protein in the pathogenesis of PD.

[0004] Antibodies targeting α-synuclein can be used to detect and analyze the expression and aggregation of α-synuclein, and can also be used to further understand the pathogenesis of Parkinson's disease and provide clues for the development of new treatments. In addition, the antibody can also be used to detect the content of α-synuclein in cerebrospinal fluid, blood or other biological samples, and can be used as a diagnostic raw material, providing an effective means for the early detection and diagnosis of Parkinson's disease. Therefore, the development and research of antibodies targeting α-synuclein has important clinical and scientific value. At present, there is still an urgent need in this field to develop a monoclonal antibody product with good affinity and specificity for α-synuclein. Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a new antibody that specifically binds to α-synuclein and its related products and uses in the art.

[0006] The present invention adopts the following technical solutions to achieve the above-mentioned invention objectives:

[0007] The first aspect of the present invention provides an anti-α-synuclein antibody.

[0008] Furthermore, HCDR1, HCDR2, and HCDR3 in the heavy chain variable region of the antibody have amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 1;

[0009] LCDR1, LCDR2, LCDR3 in the light chain variable region of the antibody have amino acid sequences such as LCDR1, LCDR2, LCDR3 in the light chain variable region shown in SEQ ID NO:2.

[0010] Furthermore, the amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region of the antibody are shown in SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, respectively;

[0011] The amino acid sequences of LCDR1, LCDR2 and LCDR3 in the light chain variable region of the antibody are shown in SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8, respectively.

[0012] Further, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO: 1 or has at least 75% homology to SEQ ID NO: 1;

[0013] Optionally, the amino acid sequence of the light chain variable region is as shown in SEQ ID NO:2 or has at least 75% homology to SEQ ID NO:2.

[0014] In some embodiments, amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence homology to the amino acid sequences corresponding to the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the present invention are also within the scope of protection of the present invention. The amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence homology include those obtained by deleting, inserting or replacing one or more amino acids or nucleotides in the parent sequence.

[0015] In some embodiments, the amino acid sequences corresponding to HCDR1, HCDR2, and HCDR3 of the present invention are not limited to the amino acid sequences shown in SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5, and the amino acid sequences corresponding to LCDR1, LCDR2, and LCDR3 of the present invention are not limited to the amino acid sequences shown in SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8. The amino acid sequences corresponding to HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 obtained by defining CDR1, CDR2, and CDR3 in the heavy chain variable region as shown in SEQ ID NO:1 and defining CDR1, CDR2, and CDR3 in the light chain variable region as shown in SEQ ID NO:2 (and the nucleotide sequences corresponding to the amino acid sequences) all fall within the protection scope of the present invention.

[0016] In some embodiments, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 can be defined by any one or any combination of two or more (two or more) of the IMGT numbering scheme, the Chothia numbering scheme, the Kabat numbering scheme, the Martin (enhanced Chothia) numbering scheme, the AbM numbering scheme, and the Aho numbering scheme (or other existing CDR numbering schemes or new CDR numbering schemes that may be generated in the future) to define the amino acid sequences corresponding to the heavy chain variable regions and light chain variable regions of the antibodies of the present invention as described above. The sequences corresponding to the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 defined in the above-mentioned manner also fall within the protection scope of the present invention.

[0017] In some embodiments, the functional variants of the antibodies provided in the first aspect of the present invention are also included in the protection scope of the present invention.

[0018] In some embodiments, the functional variant refers to a protein having a significant or significant sequence homology or similarity compared to a parent antibody (the antibody described in the first aspect of the present invention), and the functional variant retains the biological activity of the parent antibody. Functional variants encompass, for example, the following variants of an antibody (parent antibody) described herein, which retain the ability to recognize target cells to a similar extent, to the same extent, or to a higher extent compared to the parent antibody. With reference to the parent antibody, the functional variant may, for example, have at least about 30%, 50%, 70%, 75%, 80%, 85%, 90%, 95% or more homology in amino acid sequence with the parent antibody.

[0019] In some embodiments, the functional variant may, for example, comprise an amino acid sequence of a parent antibody having at least one conservative amino acid substitution. As an alternative or in addition, the functional variant may comprise an amino acid sequence of a parent antibody having at least one non-conservative amino acid substitution. In this case, the non-conservative amino acid substitution preferably does not interfere with or inhibit the biological activity of the functional variant. The non-conservative amino acid substitution can enhance the biological activity of the functional variant, so that the biological activity of the functional variant is improved compared to the parent antibody. Such functional variants obtained by conservative amino acid substitution or non-conservative amino acid substitution on the basis of the parent antibody also fall within the scope of protection of the present invention.

[0020] In some embodiments, conservative amino acid substitutions are known in the art and include replacing one amino acid with particular physical and / or chemical properties with another amino acid with the same or similar chemical or physical properties. For example, conservative amino acid substitutions can be substitutions of an acidic / negatively charged polar amino acid for another acidic / negatively charged polar amino acid (e.g., Asp or Glu), substitutions of an amino acid with a non-polar side chain for another amino acid with a non-polar side chain (e.g., Ala, Gly, Val, He, Leu, Met, Phe, Pro, Trp, Cys, Val, etc.), substitutions of a basic / positively charged polar amino acid for another basic / positively charged polar amino acid (e.g., Lys, His, Arg, etc.), substitutions of an uncharged amino acid with a polar side chain for another uncharged amino acid with a polar side chain (e.g., Asn, Gin, Ser, Thr, Tyr, etc.), substitutions of an amino acid with a β-branched side chain for another amino acid with a β-branched side chain (e.g., He, Thr and Val), substitutions of an amino acid with an aromatic side chain for another amino acid with an aromatic side chain (e.g., His, Phe, Trp and Tyr).

[0021] A second aspect of the present invention provides a bispecific antibody.

[0022] Further, the bispecific antibody comprises the antibody described in the first aspect of the present invention;

[0023] Optionally, the bispecific antibody further comprises a second antibody that specifically binds to another antigen.

[0024] In some embodiments, the second antibody that specifically binds to other antigens is not particularly limited, and can be other antibodies targeting α-synuclein or antibodies targeting any antigen other than α-synuclein. Those skilled in the art can make routine selections based on actual needs.

[0025] In some embodiments, the bispecific antibody comprises binding specificities for two different antigens (one antigen is α-synuclein and the other antigen is any antigen other than α-synuclein). In other embodiments, the bispecific antibody comprises binding specificities for different epitopes of the same antigen (e.g., α-synuclein) or the same epitope (e.g., having different binding affinities and / or specific epitopes for the same antigen).

[0026] In some embodiments, the antigens other than α-synuclein include, but are not limited to, LRRK2 (leucine-rich repeat kinase 2), dopamine receptors (D1 and D2 receptors), and tau protein.

[0027] The third aspect of the present invention provides a nucleic acid molecule.

[0028] Furthermore, the nucleic acid molecule encodes the antibody described in the first aspect of the present invention or the bispecific antibody described in the second aspect of the present invention.

[0029] A person skilled in the art can easily adopt known methods, such as directed evolution and point mutation, to mutate the nucleotide sequence corresponding to the antibody of the present invention. As long as the antibody of the first aspect of the present invention or the bispecific antibody of the second aspect of the present invention is encoded, it is derived from the nucleotide sequence of the present invention and is equivalent to the sequence of the present invention, it also falls within the scope of protection of the present invention.

[0030] In some embodiments, the nucleic acid molecule is isolated or purified. The sequence of the nucleic acid molecule can be obtained using conventional techniques or hybridoma technology. Once the relevant sequence is obtained, the relevant sequence can be obtained in large quantities by recombinant methods. This is usually cloned into a vector, then transferred into a cell, and then the relevant sequence is isolated from the host cell after proliferation by conventional methods. In addition, the relevant sequence can also be synthesized by artificial synthesis, especially when the fragment length is short. Usually, a fragment with a longer sequence can be obtained by synthesizing multiple small fragments first and then connecting them.

[0031] The fourth aspect of the present invention provides a recombinant vector.

[0032] Further, the recombinant vector comprises the nucleic acid molecule described in the third aspect of the present invention;

[0033] Optionally, the vector is a DNA vector, an RNA vector, a plasmid, a transposon vector and / or a vector of viral origin;

[0034] Optionally, the virus-derived vector is an adenoviral vector, an adeno-associated viral vector, a retroviral vector, a lentiviral vector, a poxvirus vector and / or a herpesvirus vector.

[0035] In some embodiments, the recombinant vector is an expression vector or a cloning vector.

[0036] In some embodiments, any vector capable of delivering nucleic acid is suitable for use in the present invention. In some embodiments, the vector is a viral vector. In some embodiments, the vector is a retroviral vector, a DNA vector, a murine leukemia virus vector, a SFG vector, a plasmid, an RNA vector, an adenoviral vector, a baculoviral vector, an Epstein Barr virus vector, a papovavirus vector, a vaccinia virus vector, a herpes simplex virus vector, an adenovirus-associated vector (AAV), a lentiviral vector, or any combination thereof.

[0037] A fifth aspect of the present invention provides a recombinant host cell.

[0038] Further, the recombinant host cell comprises the recombinant vector described in the fourth aspect of the present invention;

[0039] Optionally, the host cell is a mammalian cell, an insect cell and / or a yeast cell.

[0040] In some embodiments, the host cell is a eukaryotic cell. In some embodiments, the eukaryotic cell includes, but is not limited to, a mammalian cell, an insect cell, a plant cell, a yeast cell. In some embodiments, the host cell for expressing the antibody or bispecific antibody of the present invention includes Escherichia coli, a yeast cell, an insect cell, a COS cell, a CHO cell, an immune cell, etc. In some embodiments, the immune cell includes a T cell, a B cell, a NK cell, an iNKT cell, a CTL cell, a dendritic cell, a myeloid cell, a monocyte, a macrophage, or any combination thereof.

[0041] In some embodiments, the recombinant host cell is prepared by the following method: introducing the nucleic acid molecule or expression vector of the present invention as described above into the host cell, and the method of introduction includes but is not limited to: physical method, chemical method, biological method. The physical method includes but is not limited to: calcium phosphate precipitation, lipid transfection, particle bombardment, microinjection, electroporation; the chemical method includes but is not limited to: colloidal dispersion system, lipid-based system; the colloidal dispersion system includes but is not limited to: macromolecular complex, nanocapsule, microsphere, bead; the lipid-based system includes but is not limited to: water-in-oil emulsion, micelle, mixed micelle, liposome; the biological method includes but is not limited to: DNA vector, lentiviral vector, poxvirus vector, herpes simplex virus vector, adenovirus vector, adeno-associated virus vector.

[0042] In some embodiments, the nucleic acid molecules or expression vectors of the present invention as described above can be introduced into host cells by various suitable means, not limited to the methods listed in the present invention, such as calcium phosphate transfection, DEAE-dextran-mediated transfection, microinjection, electroporation, TALEN method, ZFN method, non-viral vector-mediated transfection (such as liposomes) or viral vector-mediated transfection (such as lentiviral infection, retroviral infection, adenoviral infection), and other physical, chemical or biological means for transfer into cells, such as transposon technology, CRISPR-Cas9 and other technologies.

[0043] A sixth aspect of the present invention provides any of the following products:

[0044] (1) An antibody conjugate, wherein the antibody conjugate is a complex formed by directly or indirectly coupling the antibody according to the first aspect of the present invention or the bispecific antibody according to the second aspect of the present invention to a detectable marker;

[0045] (2) A detection reagent, comprising the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, or the antibody conjugate described above;

[0046] (3) A detection product, comprising the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the antibody conjugate or the detection reagent;

[0047] (4) A pharmaceutical composition comprising the antibody according to the first aspect of the present invention or the bispecific antibody according to the second aspect of the present invention;

[0048] (5) A biological preparation comprising the pharmaceutical composition;

[0049] Optionally, the detectable label is a bioluminescent agent, a paramagnetic ion, an enzyme, a photosensitive diagnostic agent, a radionuclide, or a chemiluminescent agent;

[0050] Optionally, the detection product is a detection kit, a detection test strip or a detection chip.

[0051] In some embodiments, the bioluminescent agent includes but is not limited to: luciferin, luciferase, aequorin; the paramagnetic ions include but are not limited to: chromium (III), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), manganese (II), iron (III), iron (II), cobalt (II), gadolinium (III), vanadium (II); the enzymes include but are not limited to: horseradish peroxidase, catalase, alkaline phosphatase, urease, glucose oxidase, β-D-galactosidase, or glucoamylase; the photosensitive diagnostic agents include but are not limited to: dihydroxysilicon phthalocyanine, protoporphyrin, hematoporphyrin, methylene blue, photoporphyrin; the radionuclides include but are not limited to: 68 Ga, 86 Y. 90 Y. 89 Zr, 110 In, 111 In, 177 Lu, 18 F. 52 Fe, 62 Cu, 64 Cu, 31 I. 154-158 Gd, 32 F. 11 C. 67 Cu, 67 Ga, 94m Tc, 94 Tc, 99m Tc, 120 I. 123 I. 124 I. 125 I. 113 N. 15 O. 186 Re, 188 Re; The chemiluminescent agent includes, but is not limited to, aromatic acridinium esters, imidazoles, luminol, isoluminol, acridinium salts, and oxalates.

[0052] In some embodiments, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier and / or excipient, which are described in detail in Remington's Pharmaceutical Sciences (19th ed, 1995).

[0053] In some embodiments, the dosage form of the biologic includes, but is not limited to, solutions, emulsions, tablets, pills, powders, granules, capsules, suspensions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, or suppositories.

[0054] In some embodiments, the administration of the pharmaceutical composition or biologic includes, but is not limited to, intravenous, intramuscular, subcutaneous, intradermal, intraperitoneal, oral, topical, intranasal, intrapulmonary, rectal, etc. When administered orally, a coating may be formulated to protect the active ingredient in the pharmaceutical composition or biologic to prevent degradation in the stomach. In addition, the active ingredient may be administered by any device capable of being transferred to the target tissue.

[0055] In a specific embodiment, the pharmaceutical composition or biological preparation provided by the present invention can be prepared into various dosage forms according to actual needs, and the dosage that is beneficial to the patient can be determined by the clinician according to factors such as the type, age, weight and general disease condition of the subject, and the mode of administration. The mode of administration can be, for example, injection or any other suitable mode of administration known to those skilled in the art.

[0056] A seventh aspect of the present invention provides any of the following methods:

[0057] (1) A method for preparing the recombinant host cell according to the fifth aspect of the present invention, the method comprising: introducing the recombinant vector according to the fourth aspect of the present invention into a host cell to obtain the recombinant host cell according to the fifth aspect of the present invention;

[0058] (2) A method for producing the antibody according to the first aspect of the present invention or the bispecific antibody according to the second aspect of the present invention, the method comprising: culturing the recombinant host cell according to the fifth aspect of the present invention, and isolating the antibody or bispecific antibody from the culture product of the recombinant host cell;

[0059] (3) A method for detecting α-synuclein in a sample for non-diagnostic and non-therapeutic purposes, the method comprising: contacting the sample with the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the antibody conjugate, detection reagent or detection product described in the sixth aspect of the present invention, and detecting the formation of an immune complex between α-synuclein and the antibody;

[0060] (4) A method for inhibiting α-synuclein activity in a sample in vitro, the method comprising: contacting the sample with the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the antibody conjugate described in the sixth aspect of the present invention, a detection reagent or a detection product.

[0061] In some embodiments, methods for introducing the nucleic acid molecule into the host cell are well known to those skilled in the art, including but not limited to: physical methods, chemical methods or biological methods.

[0062] In some embodiments, the sample or the sample to be tested can be selected from blood, serum, plasma, brain tissue, cerebrospinal fluid, urine, saliva, ascites, non-tissue-associated cells, tissues, histological preparations, etc. from the subject to be tested. The present invention has no particular limitation on the specific type of the sample or the sample to be tested, and any sample that may contain α-synuclein can be used as the sample or the sample to be tested.

[0063] An eighth aspect of the present invention provides any of the following applications:

[0064] (1) Use of the antibody according to the first aspect of the present invention, the bispecific antibody according to the second aspect of the present invention, the nucleic acid molecule according to the third aspect of the present invention, the recombinant vector according to the fourth aspect of the present invention, or the recombinant host cell according to the fifth aspect of the present invention in the preparation of an antibody conjugate for detecting α-synuclein;

[0065] (2) Use of the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the nucleic acid molecule described in the third aspect of the present invention, the recombinant vector described in the fourth aspect of the present invention, the recombinant host cell described in the fifth aspect of the present invention, or the antibody conjugate described in the sixth aspect of the present invention in the preparation of a detection reagent for detecting α-synuclein;

[0066] (3) Use of the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the nucleic acid molecule described in the third aspect of the present invention, the recombinant vector described in the fourth aspect of the present invention, the recombinant host cell described in the fifth aspect of the present invention, the antibody conjugate or the detection reagent described in the sixth aspect of the present invention in the preparation of a detection product for detecting α-synuclein;

[0067] (4) Use of the antibody according to the first aspect of the present invention, the bispecific antibody according to the second aspect of the present invention, the nucleic acid molecule according to the third aspect of the present invention, the recombinant vector according to the fourth aspect of the present invention, the recombinant host cell according to the fifth aspect of the present invention, the antibody conjugate, detection reagent or detection product according to the sixth aspect of the present invention in detecting α-synuclein for non-diagnostic and non-therapeutic purposes;

[0068] (5) Use of the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the nucleic acid molecule described in the third aspect of the present invention, the recombinant vector described in the fourth aspect of the present invention, the recombinant host cell described in the fifth aspect of the present invention, the antibody conjugate, detection reagent or detection product described in the sixth aspect of the present invention in the preparation of a diagnostic product for diagnosing or assisting in the diagnosis of α-synuclein disease;

[0069] (6) Use of the antibody according to the first aspect of the present invention, the bispecific antibody according to the second aspect of the present invention, the nucleic acid molecule according to the third aspect of the present invention, the recombinant vector according to the fourth aspect of the present invention or the recombinant host cell according to the fifth aspect of the present invention in the preparation of a drug for treating and / or preventing α-synuclein disease;

[0070] (7) Use of the antibody according to the first aspect of the present invention, the bispecific antibody according to the second aspect of the present invention, the nucleic acid molecule according to the third aspect of the present invention, the recombinant vector according to the fourth aspect of the present invention, the recombinant host cell according to the fifth aspect of the present invention or the pharmaceutical composition according to the sixth aspect of the present invention in the preparation of a biological agent for treating and / or preventing α-synuclein disease;

[0071] Optionally, the α-synuclein disease is a neurodegenerative disease;

[0072] Optionally, the neurodegenerative disease is Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Lewy body variant of Alzheimer's disease, diffuse Lewy body disease, combined Alzheimer's disease and Parkinson's disease, or neurodegeneration with type I brain iron accumulation;

[0073] Optionally, the neurodegenerative disease is Parkinson's disease.

[0074] In some embodiments, the α-synuclein disease is a general term for a class of neurodegenerative diseases. Any disease caused by abnormal aggregation and deposition of α-synuclein is within the scope of protection of the present invention and is not limited to the specific disease types listed above.

[0075] The present invention provides a method for diagnosing and / or assisting in the diagnosis of alpha-synuclein disease, the method comprising: using the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the nucleic acid molecule described in the third aspect of the present invention, the recombinant vector described in the fourth aspect of the present invention, the recombinant host cell described in the fifth aspect of the present invention, the antibody conjugate, the detection reagent or the detection product described in the sixth aspect of the present invention to detect a test sample derived from a subject, and detecting the presence or absence or the content of alpha-synuclein in the test sample derived from the subject through an antigen-antibody reaction, so as to diagnose and / or assist in the diagnosis of whether the subject suffers from alpha-synuclein disease or the risk of suffering from alpha-synuclein disease.

[0076] The present invention provides a method for treating and / or preventing alpha-synuclein disease, the method comprising: administering to a subject in need thereof a therapeutically and / or preventively effective amount of the antibody described in the first aspect of the present invention, the bispecific antibody described in the second aspect of the present invention, the nucleic acid molecule described in the third aspect of the present invention, the recombinant vector described in the fourth aspect of the present invention, the recombinant host cell described in the fifth aspect of the present invention, or the pharmaceutical composition or biological preparation described in the sixth aspect of the present invention.

[0077] Compared with the prior art, the present invention has the following beneficial effects:

[0078] The present invention provides a novel monoclonal antibody targeting α-synuclein in the art, wherein the monoclonal antibody can specifically bind to α-synuclein and has good specificity and affinity. The present invention lays a foundation for the development of α-synuclein-related detection and / or auxiliary detection products in the art. At the same time, the present invention is expected to provide more diagnostic tools and treatment strategies for the diagnosis and / or treatment of α-synuclein diseases, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] Figure 1 : Affinity test result diagram of the antibody targeting α-synuclein prepared by the present invention;

[0080] Figure 2 : Specificity detection result diagram of the antibody targeting α-synuclein prepared by the present invention. DETAILED DESCRIPTION

[0081] The inventors of the present invention have conducted extensive and in-depth research and screened a novel monoclonal antibody targeting α-synuclein using monoclonal antibody technology. The monoclonal antibody can specifically bind to α-synuclein, has good specificity and affinity, and can be applied to the development of α-synuclein-related detection and / or auxiliary detection products.

[0082] The amino acid sequence of the heavy chain variable region (VH) of the monoclonal antibody targeting α-synuclein is shown in SEQ ID NO: 1, the amino acid sequence of the light chain variable region (VL) of the monoclonal antibody targeting α-synuclein is shown in SEQ ID NO: 2, the amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region are shown in SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, respectively, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region are shown in SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, respectively.

[0083] In order to better understand the technical solutions related to the present application, the terms involved in the present application are explained as follows. Unless otherwise specified, the following terms involved in the present application refer to the following contents.

[0084] As used herein, the terms "comprising" or "including" refer to compositions, methods, and their corresponding components, which are present in a given embodiment, but are also open ended and include unspecified elements.

[0085] As used herein, the term "antibody" refers to a polypeptide of the immunoglobulin family that can non-covalently, reversibly and specifically bind to a corresponding antigen. For example, a naturally occurring IgG antibody is a tetramer comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three domains CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises one domain CL.

[0086] Among them, the VH region and the VL region can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), which are interspersed with more conservative regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Among them, the three CDR regions of the heavy chain are represented by HCDR1, HCDR2 and HCDR3, respectively, and the three CDR regions of the light chain are represented by LCDR1, LCDR2 and LCDR3, respectively. The variable regions of the heavy and light chains contain binding domains that interact with antigens.

[0087] As used herein, the term "homology" refers to sequence similarity with a target amino acid sequence or nucleotide sequence. "Homology" includes amino acid sequences with 75% or more, or 85% or more, or 90% or more, or 95% or more homology to the amino acid sequence of the antibody provided by the present invention; nucleotide sequences with 75% or more, or 85% or more, or 90% or more, or 95% or more homology to the nucleotide sequence of the antibody provided by the present invention. Homology can be evaluated by the naked eye or computer software. When using computer software, the homology between two or more sequences can be expressed as a percentage (%), which can be used to evaluate the homology between related sequences. The homology of 75% or more can be 75%, 80%, 85%, 90% or more homology.

[0088] In some embodiments, modification of one or more amino acids in a protein generally does not affect the function of the protein. Those skilled in the art will recognize that changing a single amino acid or a small percentage of amino acids or individual additions, deletions, insertions, substitutions to an amino acid sequence are conservative modifications, wherein the changes in a protein produce proteins with similar functions. Conservative substitution tables that provide amino acids with similar functions are well known in the art. Such amino acid sequences obtained by conservative substitutions are also included in the scope of protection of the present invention.

[0089] Substitution, deletion, insertion or any combination thereof can be used to achieve the final derivative or variant. Typically, these changes are made on several amino acids to minimize the changes in the molecule, particularly the immunogenicity and specificity of the antigen-binding protein. However, larger changes can be tolerated in some cases. Amino acid substitutions are typically single bases; insertions will typically be on the order of about one to about twenty amino acid residues, although significantly larger insertions may be tolerated. The scope of deletions is about one to about twenty amino acid residues, although in some cases, deletions can be much larger.

[0090] As used herein, the term "nucleic acid molecule" may include natural, non-natural or altered nucleotides; and it may include natural, non-natural or altered internucleotide connections, such as phosphoramidate connections or phosphorothioate connections, replacing the phosphodiester present between the nucleotides of the unmodified oligonucleotide. In some embodiments, the nucleic acid does not include any insertion, deletion, inversion and / or replacement. However, in some cases for nucleic acid, it may be appropriate to include one or more insertions, deletions, inversions and / or replacements, and therefore, the nucleic acids formed by these insertions, deletions, inversions and / or replacements are also within the scope of the present invention.

[0091] As used herein, the term "expression vector" may be any suitable recombinant expression vector and may be used to transform or transfect any suitable host cell. Suitable vectors include those designed for proliferation and amplification or for expression or both, such as plasmids and viruses. Vectors may be selected from the pUC series (Fermentas Life Sciences, GlenBumie, MD), the pBluescript series (Stratagene, LaJolla, CA), the pET series (Novagen, Madison, WI), the pGEX series (Pharmacia Biotech, Uppsala, Sweden) and the pEX series (Clontech, Palo Alto, CA).

[0092] In some embodiments, the expression vector further comprises a vector of an appropriate promoter or control sequence. These expression vectors can be used to transform appropriate host cells to enable them to express proteins. The host cell can be a prokaryotic cell, such as a bacterial cell; or a lower eukaryotic cell, such as a yeast cell; or a higher eukaryotic cell, such as a mammalian cell. Representative examples include: bacterial cells such as Escherichia coli, Streptomyces; Salmonella typhimurium; fungal cells such as yeast; plant cells; insect cells such as Drosophila S2 or Sf9; animal cells such as CHO, COS7, NSO or Bowes melanoma cells, etc. Host cells particularly suitable for the present invention are eukaryotic host cells, especially mammalian cells, such as CHO cells, 293 cells, etc.

[0093] As used herein, the term "cloning vector" refers to a DNA molecule, such as a plasmid, cosmid or bacteriophage, that has the ability to replicate autonomously in a host cell. Cloning vectors typically contain one or a small number of restriction endonuclease recognition sites, where exogenous DNA sequences are inserted in a defined manner without losing the necessary biological functions of the vector, and marker genes, which are suitable for identification and selection of cells transformed with the cloning vector. Marker genes typically include genes that provide tetracycline resistance or ampicillin resistance.

[0094] In some embodiments, the present invention is not particularly limited to the specific type of the vector, and its selection depends on the desired function. Non-limiting examples of vectors include, but are not limited to, plasmid vectors, viral-derived vectors (e.g., lentiviral vectors, retroviral vectors, adenoviral vectors, adeno-associated viral vectors, poxvirus vectors, herpesvirus vectors, baculovirus vectors, papillomavirus vectors, papillomas), phage vectors, and other vectors conventionally used in, for example, genetic engineering. Various plasmids and vectors can be constructed based on methods well known to those skilled in the art.

[0095] In some embodiments, the vector of the present invention may contain one or more replication origins (ori) and genetic systems for cloning or expression, one or more markers (e.g., antibiotic resistance) for selection in a host, and one or more expression cassettes. In addition, the coding sequence contained in the vector can be connected to a transcriptional regulatory element and / or to other amino acid coding sequences using established methods. Such regulatory sequences are well known to those skilled in the art and include, but are not limited to, regulatory sequences that ensure transcription initiation, internal ribosome entry sites (IRES), and optional regulatory elements that ensure transcription termination and transcript stability. Non-limiting examples of such regulatory elements that ensure transcription initiation include promoters, translation initiation codons, enhancers, insulators, and / or regulatory elements that ensure transcription termination, which are included downstream of the nucleic acid molecule of the present invention. Further examples include Kozak sequences and intervening sequences flanking donor and acceptor sites for RNA splicing, nucleotide sequences encoding secretion signals, or signal sequences depending on the expression system used, which can guide the expressed protein to a cell compartment or culture medium. The vector may also contain additional expressible polynucleotides encoding one or more protein chaperones to promote correct protein folding.

[0096] In some embodiments, additional examples of suitable replication origins include, but are not limited to, full-length ColE1, truncated ColEI, SV40 virus, and M13 replication origins, while additional examples of suitable promoters include, but are not limited to, cytomegalovirus (CMV) promoter, SV40-promoter, RSV-promoter, lacZ promoter, tetracycline promoter / operator, chicken β-actin promoter, CAG-promoter, gai10 promoter, human elongation factor 1α-promoter, AOX1 promoter, GAL1 promoter, etc. An example of an enhancer is, for example, the SV40-enhancer. Non-limiting additional examples of regulatory elements that ensure transcription termination include SV40-polyadenylation site, tk-polyadenylation site, p-factor-independent lpp terminator, or AcMNPV polyhedral polyadenylation signal. Further non-limiting examples of selectable markers include dhfr, which confers resistance to methotrexate, npt, which confers resistance to the aminoglycosides neomycin, kanamycin and paromycin, and hygro, which confers resistance to hygromycin. Additional selection genes have been described, namely trpB, which allows cells to utilize indole instead of tryptophan; hisD, which allows cells to utilize histinol instead of histidine; mannose 6-phosphate isomerase, which allows cells to utilize mannose and ODC (ornithine decarboxylase), which confers resistance to the ornithine decarboxylase inhibitor 2-(difluoromethyl)-DL-ornithine DFMO or a deaminase from Aspergillus terreus that confers resistance to blasticidin S.

[0097] In order to facilitate the purification of the nucleic acid molecules of the present invention, a tag sequence can be inserted into the expression vector. Examples of tags include, but are not limited to, six histidine tags, myc tags, or FLAG tags. Any tag known to those skilled in the art that facilitates purification can be used in the present invention.

[0098] As used herein, the term "subject" includes humans and non-human animals. Non-human animals include all vertebrates (e.g., mammals and non-mammals) such as non-human primates (e.g., cynomolgus monkeys), sheep, dogs, cows, chickens, mice, amphibians, and reptiles. In a specific embodiment of the present invention, the subject is preferably a human.

[0099] As used herein, the term "therapeutically and / or prophylactically effective amount" refers to an amount of an antibody, bispecific antibody, pharmaceutical composition, and / or biologic described herein that is effective to inhibit, prevent, arrest, delay, or treat the symptoms of a particular disease, condition, or side effect. Such diseases, conditions, and side effects include, but are not limited to, those pathological conditions associated with abnormal aggregation and deposition of alpha-synuclein, wherein treatment or prevention includes, for example, inhibiting the activity of cells, tissues, or receptors by contacting them with an antibody, bispecific antibody, pharmaceutical composition, and / or biologic described herein. The therapeutically and / or prophylactically effective amount may vary depending on the route of administration and dosage form, the age and weight of the subject, and / or the disease or condition being treated.

[0100] The present invention is further described below in conjunction with specific embodiments, which are only used to explain the present invention and cannot be understood as limiting the present invention. It can be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and purpose of the present invention, and the scope of the present invention is defined by the claims and their equivalents.

[0101] The reagents and raw materials used in the present invention are easily obtained by those of ordinary skill in the art, and can be obtained from commercial sources unless otherwise specified. The experimental methods for which specific conditions are not specified in the present invention are usually carried out according to conventional conditions in the art or according to conditions recommended by the manufacturer. In particular, the following examples are only used to illustrate the present invention and should not limit the scope of the present invention in any way. It should be noted that the experimental conditions and results described in the following examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0102] Example 1 Preparation of antibodies targeting α-synuclein

[0103] 1. Preparation and processing of immunogens

[0104] Recombinant α-synuclein was used as the immunogen, and its purity and molecular weight were first identified by SDS-PAGE technology. To ensure that the immunogenicity was enhanced, we used ImmunoPlus technology to process the protein.

[0105] 2. Mouse immunization procedure

[0106] BALB / c mice were selected for routine immunization. After three immunization cycles, the titer of antiserum was detected by indirect ELISA technology, and mice with the highest titer were selected for subsequent experiments. At the same time, Western blotting technology was used to verify the specific recognition ability of antiserum for recombinant antigens.

[0107] 3. Isolation and preparation of spleen cells

[0108] After the selected mice were killed by cervical scissoring, the spleen was quickly removed under sterile conditions. The spleen was placed in a sterilized 90-100 mesh stainless steel mesh, 3 mL of serum-free culture medium was injected into the spleen using a syringe, and cells were obtained by repeated aspiration. Subsequently, the cells were made into a suspension and injected into a 50 mL centrifuge tube, and an appropriate amount of culture medium was added for blowing and standing. After centrifugation and counting, the spleen cells were set aside.

[0109] 4. Cell fusion operation

[0110] Mouse myeloma cells and spleen cells were mixed in a ratio of 1:5, and the supernatant was removed by centrifugation. Next, 1 mL of 40% PEG solution was added dropwise within 1 minute, while the centrifuge tube was gently rotated to promote cell fusion. Subsequently, 1 mL of serum-free culture medium was added dropwise within 60 seconds while continuously rotating, and then 20 mL of serum-free culture medium was slowly added within 5 minutes. After centrifugation, the supernatant was removed, the cells were suspended with complete culture medium, and gently mixed. Finally, the cell suspension was evenly distributed into a 96-well plate, placed in a 37°C, CO2 incubator for 24 hours, and then replaced with HAT selective culture medium.

[0111] 5. Cultivation of fused cells and antibody detection

[0112] After fusion, the cells were half-replaced within 7-10 days, and then half-replaced every 2-3 days. After 2-3 weeks of culture, hybrid cell colonies gradually formed. When the colony proliferated to 1 / 3 of the well area, the culture supernatant was taken for antibody detection. The ELISA method was used for detection. The recombinant protein was used as the antigen to coat the enzyme-labeled plate. The coating antigen concentration was set to 1 μg / mL, and 100 μL was added to each well. PBS (pH=7.4) was used as the coating buffer and placed overnight at 4°C. The next day, after washing with PBS three times, it was blocked with 1% BSA. Subsequently, the cell culture supernatant containing monoclonal antibodies was added for incubation, and the positive antiserum of mice and blank culture supernatant were used as positive and negative controls, respectively. After incubation, washing and addition of secondary antibody, the substrate was added for color development, and the absorbance value was measured using an enzyme marker to determine the antibody positivity. Finally, the cells that tested positive were further cloned and cultured, and the positive cell lines were determined after verification, and the proliferation, freezing and in vitro culture were performed.

[0113] 6. Purification and sequencing of cloned antibodies

[0114] The monoclonal antibodies in the mouse hybridoma cell culture medium were purified by conventional purification methods (first, the culture medium producing the monoclonal antibody was precipitated with half-saturated and saturated ammonium sulfate for preliminary concentration and purification; then further purified by affinity chromatography), and the purified monoclonal antibodies were sequenced (total RNA was extracted using Invitrogen's Trizol, reverse transcribed to generate cDNA, and then the heavy and light chain variable region genes were amplified by PCR using specific primers. After the PCR product was purified by electrophoresis, it was inserted into the vector by TA cloning, sequenced, and sequence analysis was performed).

[0115] 7. Experimental results

[0116] The sequence information corresponding to the screened antibodies targeting α-synuclein is shown in Table 1, the amino acid sequences of the corresponding heavy chain variable regions HCDR1-3 are shown in SEQ ID NOs: 3-5, respectively, the amino acid sequences of the corresponding light chain variable regions LCDR1-3 are shown in SEQ ID NOs: 6-8, respectively, the amino acid sequence of the corresponding heavy chain variable region VH is shown in SEQ ID NO: 1, and the amino acid sequence of the corresponding light chain variable region VL is shown in SEQ ID NO: 2.

[0117] Table 1 Sequence information of monoclonal antibodies targeting α-synuclein

[0118]

[0119] Example 2 Functional study of antibodies targeting α-synuclein

[0120] 1. Affinity detection of antibodies targeting α-synuclein

[0121] (1) Experimental methods

[0122] a. Coating antigen

[0123] Dilute α-synuclein antigen to a concentration of 1 µg / mL with phosphate buffered saline (PBS, pH=7.4). Then add 100 µL / well to the wells of the ELISA plate. Place the ELISA plate in a 4°C refrigerator overnight.

[0124] b. Washing

[0125] Take out the ELISA plate and discard the antigen solution in the well. Use PBS buffer containing Tween-20 as the washing solution, fill each well, gently shake the ELISA plate to make the washing solution fully contact the well wall, and then discard the washing solution. Repeat the above washing process 3 times. After each wash, fully discard the washing solution and pat the ELISA plate dry to ensure that there is no residual washing solution in the well.

[0126] c. Add primary antibody

[0127] The purified antibody was diluted in PBS buffer (1:1000 to 1:512000). Then, the diluted primary antibody solution was added to each well of the ELISA plate, 100 µL / well. The ELISA plate was placed in a 37°C incubator and incubated for 1 hour to allow the primary antibody to fully bind to the antigen.

[0128] d. Add secondary antibody

[0129] Wash the ELISA plate 5 times with washing solution, pat the plate dry, add 1:10,000 diluted secondary antibody solution, 100 µL / well. Place the ELISA plate in a 37℃ incubator and incubate for 30 minutes to allow the secondary antibody to fully bind to the primary antibody.

[0130] e. Add substrate for color development

[0131] Wash the ELISA plate 5 times and pat dry, add 50 μL of substrate A and B to each well, incubate at 37°C in the dark for 15 min. Add 50 μL of stop solution to each well, and measure the OD value of each well at a wavelength of 450 nm within 15 min.

[0132] The affinity of the α-synuclein antibody was evaluated by comparing the absorbance values ​​corresponding to different concentrations of the antigen.

[0133] (2) Experimental results

[0134] The results are as follows Figure 1As shown, the results showed that the absorbance value of the high-concentration antibody was 2.652, which was much larger than the negative control value of 0.084, indicating that the antibody targeting α-synuclein prepared in Example 1 of the present invention had a good affinity for the recombinant protein antigen (α-synuclein).

[0135] 2. Specific detection of antibodies targeting α-synuclein

[0136] (1) Experimental methods

[0137] Western immunoblotting was used to detect the specificity of the antibody for α-synuclein in plasma samples. A 10% SDS-PAGE gel was prepared, 10 μL was loaded into each well, and electrophoresis was performed using a PowerPac™ HC high current electrophoresis instrument. The sample was transferred to a PVDF membrane using a multi-channel semi-dry transfer instrument. The PVDF membrane was blocked with 5% skim milk powder for 30 min, and the purified α-synuclein antibody was added and incubated at room temperature for 2 h. After washing the membrane with TBST, HRP-labeled secondary antibody was added and incubated at room temperature for 1 h. The Omni-ECLTM ultrasensitive chemiluminescence detection kit was used for color development, and the image was taken on an electronic compression imager.

[0138] (2) Experimental results

[0139] The results are as follows Figure 2 As shown, when the antibody is used to detect plasma samples, a target band can be displayed at a position around 15 KD, and the band is single, indicating that the antibody targeting α-synuclein prepared in Example 1 of the present invention can specifically detect α-synuclein in the sample.

Claims

1. An anti-α-synuclein antibody, characterized in that The HCDR1, HCDR2, and HCDR3 in the heavy chain variable region of the antibody have the amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 1; LCDR1, LCDR2, LCDR3 in the light chain variable region of the antibody have amino acid sequences such as LCDR1, LCDR2, LCDR3 in the light chain variable region shown in SEQ ID NO:

2.

2. The antibody according to claim 1, characterized in that The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region of the antibody are shown in SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, respectively; The amino acid sequences of LCDR1, LCDR2 and LCDR3 in the light chain variable region of the antibody are shown in SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8, respectively.

3. The antibody according to claim 1, characterized in that The amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO: 1 or has at least 75% homology to SEQ ID NO: 1; The amino acid sequence of the light chain variable region is as shown in SEQ ID NO: 2 or has at least 75% homology to SEQ ID NO:

2.

4. A bispecific antibody, characterized in that: The bispecific antibody comprises the antibody of any one of claims 1-3.

5. A nucleic acid molecule, characterized in that The nucleic acid molecule encodes the antibody according to any one of claims 1 to 3 or the bispecific antibody according to claim 4.

6. A recombinant vector, characterized in that: The recombinant vector comprises the nucleic acid molecule of claim 5.

7. A recombinant host cell, characterized in that The recombinant host cell comprises the recombinant vector according to claim 6.

8. Any of the following products, characterized in that: The products include: (1) An antibody conjugate, wherein the antibody conjugate is a complex formed by directly or indirectly coupling the antibody according to any one of claims 1 to 3 or the bispecific antibody according to claim 4 to a detectable marker; (2) A detection reagent, comprising the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, or the antibody conjugate; (3) A detection product, comprising the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the antibody conjugate or the detection reagent; (4) A pharmaceutical composition comprising the antibody according to any one of claims 1 to 3 or the bispecific antibody according to claim 4; (5) A biological preparation comprising the pharmaceutical composition.

9. Any of the following methods, characterized in that: The method comprises: (1) A method for preparing the recombinant host cell according to claim 7, the method comprising: introducing the recombinant vector according to claim 6 into a host cell to obtain the recombinant host cell according to claim 7; (2) A method for producing the antibody according to any one of claims 1 to 3 or the bispecific antibody according to claim 4, characterized in that the method comprises: culturing the recombinant host cell according to claim 7, and isolating the antibody or bispecific antibody from the culture product of the recombinant host cell; (3) A method for detecting α-synuclein in a test sample for non-diagnostic and non-therapeutic purposes, the method comprising: contacting the test sample with the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the antibody conjugate, the detection reagent or the detection product according to claim 8, and detecting the formation of an immune complex between α-synuclein and the antibody; (4) A method for inhibiting α-synuclein activity in a sample in vitro, the method comprising: contacting the sample with the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the antibody conjugate according to claim 8, the detection reagent or the detection product.

10. Any of the following applications, characterized in that: The applications include: (1) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6 or the recombinant host cell according to claim 7 in the preparation of an antibody conjugate for detecting α-synuclein; (2) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6, the recombinant host cell according to claim 7 or the antibody conjugate according to claim 8 in the preparation of a detection reagent for detecting α-synuclein; (3) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6, the recombinant host cell according to claim 7, the antibody conjugate or the detection reagent according to claim 8 in the preparation of a detection product for detecting α-synuclein; (4) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6, the recombinant host cell according to claim 7, the antibody conjugate, the detection reagent or the detection product according to claim 8 in detecting α-synuclein for non-diagnostic and non-therapeutic purposes; (5) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6, the recombinant host cell according to claim 7, the antibody conjugate, the detection reagent or the detection product according to claim 8 in the preparation of a diagnostic product for diagnosing or assisting in the diagnosis of α-synuclein disease; (6) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6 or the recombinant host cell according to claim 7 in the preparation of a medicament for treating and / or preventing α-synuclein disease; (7) Use of the antibody according to any one of claims 1 to 3, the bispecific antibody according to claim 4, the nucleic acid molecule according to claim 5, the recombinant vector according to claim 6, the recombinant host cell according to claim 7 or the pharmaceutical composition according to claim 8 in the preparation of a biological agent for treating and / or preventing α-synuclein disease.

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