Anti-human NR1 antibody derivative

Abstract

The present invention solves the problem of providing a novel therapeutic agent for treating patients suffering from anti-NMDAR encephalitis. Patients suffering from anti-NMDAR encephalitis have pathogenic anti-human NR1 antibodies, which induce the internalization of NMDAR on the cell surface. As a result, the NMDAR function in the brains of such patients is weakened. The inventors of the present invention have found that the single-arm anti-human NR1 antibody according to the present invention competitively binds to NR1 with the pathogenic anti-human NR1 antibody and inhibits the internalization of NMDAR by the pathogenic anti-human NR1 antibody, thereby showing a therapeutic effect against anti-NMDAR encephalitis. Accordingly, the present invention provides a single-arm anti-human NR1 antibody, a polynucleotide encoding the antibody, an expression vector containing the polynucleotide, a host cell transformed by the expression vector, a method for producing the antibody, a pharmaceutical composition containing the antibody, the use of the antibody in the manufacture of the pharmaceutical composition, and a method for treating anti-NMDAR encephalitis using the antibody.

Description

Technical Field

[0001] The present invention relates to an anti-human NR1 antibody derivative used as an active ingredient in a pharmaceutical composition. Background Art

[0002] The relationship between autoantibodies against nerve cells and diseases has recently become clear. Autoantibodies against nerve cells are called antineuronal antibodies, and they are considered to be the cause of nervous system diseases such as autoimmune encephalitis. In recent years, novel antineuronal antibodies against cell surface and synaptic antigens have been successively identified. Many of them are antibodies against receptors or channels expressed on nerve cells or their related molecules (Journal of Clinical and Experimental Medicine (IGAKU NO AYUMI), 2015, Vol. 25, No. 5, pp. 497-503 / Neuro-oncol. 2014., Vol. 16, pp. 771-778).

[0003] In 2007, Dalmau et al. found that in encephalitis with a unique clinical course complicated by ovarian teratoma, the autoantibodies against cell surface antigens present in the patient's blood and cerebrospinal fluid were antibodies against the NR1 (also called GluN1) subunit of the NMDA (N-methyl-D-aspartic acid) receptor (hereinafter referred to as "NMDAR"), and this encephalitis was named "anti-NMDAR encephalitis". Among autoimmune encephalitides, anti-NMDAR encephalitis is one of the most common autoimmune encephalitides, and its clinical manifestations are clearly defined. In patients with anti-NMDAR encephalitis, mental symptoms usually appear first, followed by various neurological symptoms, including memory impairment, speech disorder, disorientation, seizures, autonomic symptoms, central hypoventilation, and involuntary movements. The pathogenesis of anti-NMDAR encephalitis progresses rapidly, and in some cases, the course of the disease is severe, requiring intensive treatment within a few weeks after the initial symptoms are found, and the treatment lasts for several months to more than one year. The anti-NR1 antibody titer in the patient's cerebrospinal fluid reflects the course of the disease. Anti-NMDAR encephalitis rarely occurs in children, the elderly, and men, but is more common in young women. It has been reported that many women with the disease are complicated by ovarian teratoma (Lancet Neurol., 2013, Vol. 12, pp. 157-165).

[0004] Pathologically, IgG deposition and reduced NMDAR expression have been reported in the brains of patients with anti-NMDAR encephalitis (Acta Neuropathol., 2009, Vol. 118, pp. 737-743, Non-Patent Document 1). On the other hand, since no deposition of cytotoxic T cells or complement has been observed in the patients' brains (Acta Neuropathol., 2009, Vol. 118, pp. 737-743), it is hypothesized that the NR1 antibody alone affects the pathogenesis. In vitro experiments have reported that the NR1 antibody derived from patients crosslinks with NMDAR and internalizes NMDAR to reduce NMDAR in the postsynaptic membrane (Non-Patent Document 1). In addition, since the symptoms of anti-NMDAR encephalitis are similar to those observed when NMDAR antagonists (ketamine and phencyclidine) are administered to healthy subjects, it is thought that the internalization of NR1 antibody in patients with NMDAR and the subsequent reduction of NMDAR clusters in the postsynaptic membrane lead to a decline in NMDAR function, thus causing the symptoms of anti-NMDAR encephalitis.

[0005] In tumor-related cases of anti-NMDAR encephalitis, early tumor resection and immunotherapy are recommended. As immunotherapy, steroid pulse therapy, plasma exchange, and immunoglobulin therapy are used. In refractory cases, cyclophosphamide pulse therapy or rituximab (which is an anti-CD20 antibody) is administered. However, due to the severity of the disease and the prolongation of the course, not all patients have a good prognosis. It has been reported that complete or near-complete recovery accounts for only 75%, while 25% remain severely disabled or dead (Lancet Neurol., 2008, Vol. 7, pp. 1091-1098).

[0006] Currently, the treatment for anti-NMDAR encephalitis does not specifically control or remove the autoantibodies that cause the pathological condition.

[0007] NMDAR is a glutamate receptor that is mainly expressed in the central nervous system and is related to memory and learning. NMDAR is an ion channel-coupled receptor that allows cations (such as sodium, potassium, and calcium ions) to permeate by binding to glutamate as a ligand. NMDAR has four subunits, consisting of two groups of heterodimers, NR1 and NR2. There are four types of NR2: NR2A, NR2B, NR2C, and NR2D. The binding site for glutamate exists in the NR2 subunit.

[0008] As anti-NR1 monoclonal antibodies, M68 (Non-Patent Document 2), MAB363 (Non-Patent Document 3), R1JHL (Non-Patent Document 4), and D65B8 (Non-Patent Document 5) have been reported. Additionally, monoclonal antibodies that bind to the N-terminal site of human NR1 and inhibit the interaction with t-PA have been reported (Patent Document 1). Furthermore, several anti-NR1 monoclonal antibodies derived from patients with anti-NMDAR encephalitis have been identified (Non-Patent Document 6, Patent Document 2). An isolated anti-NR1 antibody derived from a patient with anti-NMDAR encephalitis reduced NMDAR clustering in vitro and inhibited NMDAR function in primary neurons derived from the mouse hippocampus (Non-Patent Document 6).

[0009] On the other hand, no anti-human NR1 antibody or its derivative has been reported to show a therapeutic effect in patients with anti-NMDAR encephalitis.

[0010] Prior Art Documents

[0011] [Patent Document]

[0012] Patent Document 1: WO2014 / 187879

[0013] Patent Document 2: WO2017 / 029299

[0014] [Non-Patent Document]

[0015] Non-Patent Document 1: [Journal of Neuroscience, (J.Neurosci.)], (USA), 2010; 30(17):5866 - 5875

[0016] Non-Patent Document 2: [Science (Science)], (USA), 2016; 352(6288):982 - 986

[0017] Non-Patent Document 3: [Journal of neurodegenerative diseases (J.Neurodegener.Dis.)], (UK), 2014; Article ID: 938530

[0018] Non-Patent Document 4: [JCI Insight (JCI Insight)], (USA), 2019; 4(15):e127736

[0019] Non-Patent Document 5: [Experimental Neurology (Exp.Neurol.)], (USA), 2017; 289:31 - 45

[0020] Non-Patent Document 6: [Brain(Brain)], (UK), 2016; 139(10): 2641-2652 Summary of the Invention

[0021] Problems to be Solved by the Invention

[0022] The present invention solves the problem of providing a novel therapeutic agent for treating patients with anti-NMDAR encephalitis.

[0023] Means for Solving the Problems

[0024] The present inventors conducted extensive research on the production of a novel therapeutic agent for treating patients with anti-NMDAR encephalitis. First, the present inventors generated single-chain anti-human NR1 antibodies from the sequence information of anti-human NR1 antibody clones having high binding activity to NMDAR derived from patients with anti-NMDAR encephalitis, and found that when bound to human NMDAR in vitro, the single-chain anti-human NR1 antibodies restored the cell surface expression level and function of human NMDAR caused by pathogenic NR1 antibodies derived from patients (Examples 1-4). In addition, based on these findings, the present inventors also generated multiple single-chain anti-human NR1 antibodies, which exhibited higher binding affinity than pathogenic NR1 antibodies, and significantly restored the cell surface expression level and function of NMDAR caused by pathogenic anti-human NR1 antibodies (Examples 5-10). In addition, the present inventors found that the newly obtained single-chain anti-human NR1 antibodies significantly improved the symptoms of an animal model of anti-NMDAR encephalitis compared with control antibodies (Example 12). Thus, the present inventors completed the present invention.

[0025] According to the present invention, the following inventions are provided as substances or methods useful in medicine and industry, for example:

[0026] A single-chain anti-human NR1 antibody selected from any one of the following (1) to (5):

[0027] (1) A single-chain anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, the heavy-chain variable region comprising CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO: 2, CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO: 2, and the light-chain variable region comprising CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO: 4, CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO: 4, and CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO: 4;

[0028] (2) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:6, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:6, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO:6, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:8, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:8, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO:8;

[0029] (3) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:10, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:10, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO:10, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:12, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:12, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:12;

[0030] (4) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:14, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:14, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:14, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:16, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:16, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:16;

[0031] (5) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20.

[0032] The single-arm anti-human NR1 antibody according to [1], selected from any one of the following (1) to (5):

[0033] (1) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4;

[0034] (2) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0035] (3) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0036] (4) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0037] (5) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:18, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:20.

[0038] The single-arm anti-human NR1 antibody according to [1], selected from any one of the following (1) to (5):

[0039] (1) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, a CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and a CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and a CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4;

[0040] (2) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:6, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:6, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO:6; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:8, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:8, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO:8;

[0041] (3) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO:10; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:12;

[0042] (4) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises a CDR1 consisting of the amino - acid sequence from amino - acid number 31 to 36 of SEQ ID NO:14, a CDR2 consisting of the amino - acid sequence from amino - acid number 51 to 66 of SEQ ID NO:14, and a CDR3 consisting of the amino - acid sequence from amino - acid number 99 to 108 of SEQ ID NO:14; the light chain comprises a light - chain variable region, and the light - chain variable region comprises a CDR1 consisting of the amino - acid sequence from amino - acid number 24 to 34 of SEQ ID NO:16, a CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 56 of SEQ ID NO:16, and a CDR3 consisting of the amino - acid sequence from amino - acid number 89 to 97 of SEQ ID NO:16;

[0043] (5) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises a CDR1 consisting of the amino - acid sequence from amino - acid number 31 to 36 of SEQ ID NO:18, a CDR2 consisting of the amino - acid sequence from amino - acid number 51 to 66 of SEQ ID NO:18, and a CDR3 consisting of the amino - acid sequence from amino - acid number 99 to 108 of SEQ ID NO:18; the light chain comprises a light - chain variable region, and the light - chain variable region comprises a CDR1 consisting of the amino - acid sequence from amino - acid number 24 to 34 of SEQ ID NO:20, a CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 56 of SEQ ID NO:20, and a CDR3 consisting of the amino - acid sequence from amino - acid number 89 to 97 of SEQ ID NO:20.

[0044] The single - arm anti - human NR1 antibody according to [3], selected from any one of the following (1) to (5):

[0045] (1) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region is composed of the amino - acid sequence shown in SEQ ID NO:2; the light chain comprises a light - chain variable region, and the light - chain variable region is composed of the amino - acid sequence shown in SEQ ID NO:4;

[0046] (2) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region is composed of the amino - acid sequence shown in SEQ ID NO:6; the light chain comprises a light - chain variable region, and the light - chain variable region is composed of the amino - acid sequence shown in SEQ ID NO:8;

[0047] (3) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10, the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 12;

[0048] (4) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 14, the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 16;

[0049] (5) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 18, the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 20.

[0050] The single-arm anti-human NR1 antibody according to any one of [1] to [4], which comprises an Fc region, and the Fc region comprises LALA mutations (L234A and L235A).

[0051] The single-arm anti-human NR1 antibody according to any one of [1] to [4], which comprises an Fc region, and the Fc region comprises a mortise and tenon mutation.

[0052] The single-arm anti-human NR1 antibody according to any one of [1] to [4], which comprises an Fc region, and the Fc region comprises LALA mutations (L234A and L235A) and a mortise and tenon mutation.

[0053] The single-arm anti-human NR1 antibody according to [6] or [7], wherein the mortise and tenon mutation is a T366W mutation in one Fc polypeptide forming the Fc region, and T366S, L368A, and Y407V mutations in another Fc polypeptide forming the Fc region.

[0054] The single-arm anti-human NR1 antibody according to any one of [1] to [8], wherein the single-arm anti-human NR1 antibody is a human antibody or a humanized antibody.

[0055] A single-arm anti-human NR1 antibody, which comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 24, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0056] A single - arm anti - human NR1 antibody, which comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26, a light chain consisting of the amino acid sequence shown in SEQ ID NO:28, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0057] A single - arm anti - human NR1 antibody, which comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30, a light chain consisting of the amino acid sequence shown in SEQ ID NO:32, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0058] A single - arm anti - human NR1 antibody, which comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:34, a light chain consisting of the amino acid sequence shown in SEQ ID NO:36, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0059] A single - arm anti - human NR1 antibody, which comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:38, a light chain consisting of the amino acid sequence shown in SEQ ID NO:40, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0060] A single - arm anti - human NR1 antibody, wherein the single - arm anti - human NR1 antibody competes with an anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4 for binding to human NR1, and inhibits NMDAR endocytosis induced by pathogenic anti - human NR1 antibodies derived from patients with anti - NMDAR encephalitis.

[0061] The single - arm anti - human NR1 antibody according to any one of [1] to

[15] , wherein the single - arm anti - human NR1 antibody is post - translationally modified.

[0062] The single - arm anti - human NR1 antibody according to

[16] , wherein the post - translational modification is pyroglutamylation of the N - terminal heavy - chain variable region and / or deletion of the C - terminal heavy - chain lysine.

[0063] A polynucleotide comprising the nucleotide sequences listed in the following (a) and / or (b):

[0064] (a) A nucleotide sequence encoding the heavy - chain variable region of the single - arm anti - human NR1 antibody according to any one of (2) to (5) of [1] and (2) to (5) of [2];

[0065] (b) The nucleotide sequence encoding the light chain variable region of the single-chain anti-human NR1 antibody as described in any one of (2) to (5) of [1] and (2) to (5) of [2].

[0066] A polynucleotide comprising the nucleotide sequence listed in (a) and / or (b) below:

[0067] (a) The nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody as described in any one of (2) to (5) of [3] and (2) to (5) of [4];

[0068] (b) The nucleotide sequence encoding the light chain of the single-chain anti-human NR1 antibody as described in any one of (2) to (5) of [3] and (2) to (5) of [4].

[0069] A polynucleotide comprising the nucleotide sequence listed in (a) and / or (b) below:

[0070] (a) The nucleotide sequence encoding the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26;

[0071] (b) The nucleotide sequence encoding the light chain consisting of the amino acid sequence shown in SEQ ID NO:28.

[0072] A polynucleotide comprising the nucleotide sequence listed in (a) and / or (b) below:

[0073] (a) The nucleotide sequence encoding the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30;

[0074] (b) The nucleotide sequence encoding the light chain consisting of the amino acid sequence shown in SEQ ID NO:32.

[0075] A polynucleotide comprising the nucleotide sequence listed in (a) and / or (b) below:

[0076] (a) The nucleotide sequence encoding the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:34;

[0077] (b) The nucleotide sequence encoding the light chain consisting of the amino acid sequence shown in SEQ ID NO:36.

[0078] A polynucleotide comprising the nucleotide sequence listed in (a) and / or (b) below:

[0079] (a) The nucleotide sequence encoding the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:38;

[0080] (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:40.

[0081] A polynucleotide comprising the nucleotide sequence listed in (a) and / or (b) below:

[0082] (a) A nucleotide sequence encoding the heavy chain of a single-chain anti-human NR1 antibody as described in

[15] ;

[0083] (b) A nucleotide sequence encoding the light chain of a single-chain anti-human NR1 antibody as described in

[15] .

[0084] An expression vector comprising a polynucleotide that comprises the nucleotide sequence listed in (a) and / or (b) below:

[0085] (a) A polynucleotide comprising a nucleotide sequence encoding the heavy chain variable region of a single-chain anti-human NR1 antibody listed in any one of (2) to (5) of [1] and (2) to (5) of [2];

[0086] (b) A polynucleotide comprising a nucleotide sequence encoding the light chain variable region of a single-chain anti-human NR1 antibody listed in any one of (2) to (5) of [1] and (2) to (5) of [2].

[0087] An expression vector comprising a polynucleotide that comprises the nucleotide sequence listed in (a) and / or (b) below:

[0088] (a) A polynucleotide comprising a nucleotide sequence encoding the heavy chain of a single-chain anti-human NR1 antibody listed in any one of (2) to (5) of [3] and (2) to (5) of [4];

[0089] (b) A polynucleotide comprising a nucleotide sequence encoding the light chain of a single-chain anti-human NR1 antibody listed in any one of (2) to (5) of [3] and (2) to (5) of [4].

[0090] The expression vector as described in

[25] or

[26] , which further comprises a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide.

[0091] An expression vector that comprises the polynucleotide listed in (a) and / or (b) below, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42:

[0092] (a) A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22;

[0093] (b) A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24.

[0094] An expression vector comprising the polynucleotide(s) listed in (a) and / or (b) below:

[0095] (a) A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26;

[0096] (b) A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28.

[0097] An expression vector comprising the polynucleotide(s) listed in (a) and / or (b) below:

[0098] (a) A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30;

[0099] (b) A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32.

[0100] An expression vector comprising the polynucleotide(s) listed in (a) and / or (b) below:

[0101] (a) A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:34;

[0102] (b) A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:36.

[0103] An expression vector comprising the polynucleotide(s) listed in (a) and / or (b) below:

[0104] (a) A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:38;

[0105] (b) A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:40.

[0106] The expression vector according to any one of

[29] to

[32] , further comprising a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0107] An expression vector, which comprises a polynucleotide containing a nucleotide sequence as listed in the following (a) to (f):

[0108] (a) A polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-chain anti-human NR1 antibody as described in

[15] ;

[0109] (b) A polynucleotide containing a nucleotide sequence encoding the light chain of a single-chain anti-human NR1 antibody as described in

[15] ;

[0110] (c) A polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-chain anti-human NR1 antibody as described in

[15] , and a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody;

[0111] (d) A polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-chain anti-human NR1 antibody as described in

[15] , and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of said antibody;

[0112] (e) A polynucleotide containing a nucleotide sequence encoding the light chain of a single-chain anti-human NR1 antibody as described in

[15] , and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of said antibody;

[0113] (f) A polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-chain anti-human NR1 antibody as described in

[15] , a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody, and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of said antibody.

[0114] A host cell transformed with one or more expression vectors selected from the following (a) to (d):

[0115] (a) An expression vector containing a polynucleotide containing a nucleotide sequence encoding the heavy chain variable region of a single-chain anti-human NR1 antibody as listed in any one of (2) to (5) of [1] and (2) to (5) of [2];

[0116] (b) An expression vector containing a polynucleotide containing a nucleotide sequence encoding the light chain variable region of a single-chain anti-human NR1 antibody as listed in any one of (2) to (5) of [1] and (2) to (5) of [2];

[0117] (c) An expression vector containing a polynucleotide containing a nucleotide sequence encoding the heavy chain variable region of a single-chain anti-human NR1 antibody as listed in any one of (2) to (5) of [1] and (2) to (5) of [2], and a polynucleotide containing a nucleotide sequence encoding the light chain variable region of said antibody;

[0118] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain variable region of a single-arm anti-human NR1 antibody listed in any one of (2) to (5) of [1] and (2) to (5) of [2], and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain variable region of said antibody.

[0119] A host cell transformed with one or more expression vectors selected from the following (a) to (d):

[0120] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (2) to (5) of [3] and (2) to (5) of [4];

[0121] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of a single-arm anti-human NR1 antibody listed in any one of (2) to (5) of [3] and (2) to (5) of [4];

[0122] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (2) to (5) of [3] and (2) to (5) of [4], and a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody;

[0123] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (2) to (5) of [3] and (2) to (5) of [4], and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody.

[0124] A host cell transformed with one or more expression vectors selected from the following (a) to (i):

[0125] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of said antibody;

[0126] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of said antibody;

[0127] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0128] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0129] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0130] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0131] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0132] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0133] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody listed in any one of (1) to (5) of [3] and (1) to (5) of [4], a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody.

[0134] A host cell transformed with one or more expression vectors selected from the following (a) to (i):

[0135] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0136] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0137] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0138] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0139] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0140] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24;

[0141] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0142] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0143] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0144] A host cell transformed with one or more expression vectors selected from the following (a) to (m):

[0145] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26;

[0146] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28;

[0147] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28;

[0148] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28;

[0149] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0150] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0151] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0152] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0153] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0154] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28;

[0155] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0156] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0157] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0158] A host cell transformed with one or more expression vectors selected from the following (a) to (m):

[0159] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30;

[0160] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0161] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0162] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0163] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0164] (f) An expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32 and a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0165] (g) An expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30, and an expression vector comprising a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0166] (h) An expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32, and an expression vector comprising a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0167] (i) An expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30 and a polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32, and an expression vector comprising a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0168] (j) An expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30 and a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42, and an expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32;

[0169] (k) An expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30, and an expression vector comprising a polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32 and a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0170] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0171] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0172] A host cell transformed with one or more expression vectors selected from the following (a) to (m):

[0173] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34;

[0174] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0175] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0176] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0177] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0178] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0179] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0180] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0181] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0182] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0183] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0184] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0185] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0186] A host cell transformed with one or more expression vectors selected from the following (a) to (m):

[0187] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38;

[0188] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0189] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0190] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0191] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0192] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0193] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0194] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0195] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0196] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0197] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0198] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0199] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0200] A host cell transformed with one or more expression vectors selected from the following (a) to (m):

[0201] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of the single-arm anti-human NR1 antibody as described in

[15] ;

[0202] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of the single-arm anti-human NR1 antibody as described in

[15] ;

[0203] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of the single-arm anti-human NR1 antibody as described in

[15] and a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody;

[0204] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of the single-arm anti-human NR1 antibody as described in

[15] , and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody;

[0205] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of the single-arm anti-human NR1 antibody as described in

[15] and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of the antibody;

[0206] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of the single-arm anti-human NR1 antibody as described in

[15] and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of the antibody;

[0207] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody as described in

[15] , and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0208] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of a single-arm anti-human NR1 antibody as described in

[15] , and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0209] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody and a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0210] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0211] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0212] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0213] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody, a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody.

[0214] A method for producing a single-arm anti-human NR1 antibody, which comprises culturing a host cell as described in any one of

[35] to

[43] and expressing the polypeptides constituting the single-arm anti-human NR1 antibody.

[0215] A pharmaceutical composition comprising a single-arm anti-human NR1 antibody as described in any one of [1] to

[17] and a pharmaceutically acceptable excipient.

[0216] The pharmaceutical composition as described in

[45] , wherein the pharmaceutical composition is used for treating anti-NMDAR encephalitis.

[0217] A method for treating anti-NMDAR encephalitis, which comprises administering a therapeutically effective amount of a single-arm anti-human NR1 antibody as described in any one of [1] to

[17] .

[0218] The single-arm anti-human NR1 antibody as described in any one of [1] to

[17] , which is used for treating anti-NMDAR encephalitis.

[0219] Use of the single-arm anti-human NR1 antibody as described in any one of [1] to

[17] in producing a pharmaceutical composition for treating anti-NMDAR encephalitis.

[0220] A monovalent anti-human NR1 antibody derivative selected from any one of the following (1) to (4):

[0221] (1) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 6, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6, and the light chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8;

[0222] (2) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10, and the light chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12;

[0223] (3) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:14, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:14, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:14, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:16, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:16, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:16;

[0224] (4) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20.

[0225] A monovalent anti-human NR1 antibody derivative selected from any one of the following (1) to (4):

[0226] (1) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6, and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0227] (2) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10, and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0228] (3) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14, and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0229] (4) A monovalent anti-human NR1 antibody derivative, which comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 18, and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 20.

[0230] A monovalent anti-human NR1 antibody derivative, wherein the antibody competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 4, and inhibits NMDAR endocytosis induced by pathogenic anti-human NR1 antibodies derived from patients with anti-NMDAR encephalitis.

[0231] A polypeptide, which comprises a nucleotide sequence encoding the monovalent anti-human NR1 antibody derivative as described in any one of

[50] to

[52] .

[0232] An expression vector, which comprises the polynucleotide as described in

[53] .

[0233] A host cell, which is transformed with the expression vector as described in

[54] .

[0234] A method for producing a monovalent anti-human NR1 antibody derivative, which comprises culturing the host cell as described in

[55] , and expressing the polypeptide constituting the monovalent anti-human NR1 antibody derivative.

[0235] A pharmaceutical composition, which comprises the monovalent anti-human NR1 antibody derivative as described in any one of

[50] to

[52] and a pharmaceutically acceptable excipient.

[0236] The pharmaceutical composition as described in

[57] , wherein the pharmaceutical composition is used for treating anti-NMDAR encephalitis.

[0237] A method for treating anti-NMDAR encephalitis, which comprises administering a therapeutically effective amount of the monovalent anti-human NR1 antibody derivative as described in any one of

[50] to

[52] .

[0238] The monovalent anti-human NR1 antibody derivative as described in any one of

[50] to

[52] , which is used for treating anti-NMDAR encephalitis.

[0239] Use of the monovalent anti-human NR1 antibody derivative as described in any one of

[50] to

[52] in the production of a pharmaceutical composition for treating anti-NMDAR encephalitis.

[0240] Effects of the present invention

[0241] The monovalent human NR1 antibody derivative of the present invention can be used as a therapeutic agent by competitively binding to NMDAR with pathogenic anti-human NR1 antibodies derived from anti-NMDAR encephalitis patients and inhibiting NMDAR internalization. BRIEF DESCRIPTION OF THE DRAWINGS

[0242] Figure 1 Shows the binding activities of the pathogenic anti-human NR1 antibody P003-102 derived from P003-102 and the single-arm anti-human NR1 antibody (ABN-5) to HEK cells expressing NMDAR. The vertical axis represents the binding activity (ΔGeoMFI: the difference between the geometric mean fluorescence intensity when the test antibody is added and the geometric mean fluorescence intensity when the test antibody is not added (only the fluorescently labeled secondary antibody)), and the horizontal axis represents the antibody concentration.

[0243] Figure 2 Shows the inhibitory activity of the novel mouse anti-human NR1 antibody isolated in Example 5 against the binding of the pathogenic anti-human NR1 antibody P003-102 to human NR1. The vertical axis represents the inhibitory activity (inhibition rate) of the mouse anti-human NR1 antibody against the binding of biotin-labeled P003-102 to human NR1 protein. The horizontal axis represents the antibody concentration.

[0244] Figure 3 Shows the binding activity of the humanized single-arm anti-human NR1 antibody to HEK cells expressing NMDAR. The vertical axis represents the geometric mean fluorescence intensity (GeoMFI), and the horizontal axis represents the antibody concentration.

[0245] Figure 4 Shows the inhibitory activities of ABN-2, ABN-4, and ABN-5 against the pathogenic anti-human NR1 antibody P003-102. The vertical axis represents the inhibitory activity (inhibition rate) of biotin-labeled P003-102 against the binding to human NR1 protein, and the horizontal axis represents the antibody concentration.

[0246] Figure 5 Shows the rescue effect of ABN-2 on the NMDAR expression level on the cell surface caused by the pathogenic anti-human NR1 antibody mixture. The vertical axis represents the NR1 expression rate (%) on the cell surface of HEK cells expressing NMDAR, and the horizontal axis represents the antibody concentration.

[0247] Figure 6 Shows the rescue effect of ABN-2 on the decrease in NMDAR ion channel function caused by the pathogenic anti-human NR1 antibody P003-102. The vertical axis represents the calcium ion influx induced by NMDA stimulation, and the horizontal axis represents the antibody concentration.

[0248] Figure 7Shows the therapeutic effect of ABN-2 on common marmosets (hereinafter referred to as "marmosets"), in which anti-NMDAR encephalitis-like symptoms were induced by continuous intraventricular administration of the pathogenic anti-human NR1 antibody P003-102. Day 0 is the day when P003-102 was first administered to the marmosets in the control antibody administration group (P003-102 + control antibody, n = 3) and the ABN-2 administration group (P003-102 + ABN-2, n = 9). From day 14 (day 14) to day 28 (day 28), the control antibody or ABN-2 was continuously administered intraventricularly for 14 days. Figure 7 The left, middle, and right vertical axes respectively represent the Abnormal Rating Scale (ARS), which was modified from the Unified Parkinson's Disease Rating Scale (UPDRS, Behav. Brain Res. 2008, Vol. 194, pp. 152-161). Respectively, Figure 7 The left side of shows the ARS of the control antibody administration group and the ABN-2 administration group on day 0, Figure 7 The middle of shows the ARS on day 14, and Figure 7 The right side of shows the ARS on day 28.

[0249] Figure 8 Shows the therapeutic effect of ABN-2 on common marmosets, where for each individual in the ABN-2 administration group (P003-102 + ABN-2), anti-NMDAR encephalitis-like symptoms were induced by continuous intraventricular administration of the pathogenic anti-human NR1 antibody P003-102, as the change of ARS over time.

[0250] Figure 9 Shows a schematic diagram of a single-arm antibody. Detailed implementation

[0251] The following is a detailed description of the present invention.

[0252] Antibody (or immunoglobulin) molecules are glycoproteins composed of multiple units. There are five classes of antibodies: IgG, IgM, IgA, IgD, and IgE. The basic structure of antibody molecules common to all classes is a Y-shaped four-chain structure, which includes two heavy chains with a molecular weight of 50,000 to 70,000 and two light chains with a molecular weight of 20,000 to 30,000. Heavy chains are usually composed of polypeptide chains containing approximately 440 amino acids and have characteristic structures according to the class, and they are called Igγ, Igμ, Igα, Igδ, and Igε, corresponding to IgG, IgM, IgA, IgD, and IgE, respectively. In addition, IgG has subclasses of IgG1, IgG2, IgG3, and IgG4, and the corresponding heavy chains are called Igγ1, Igγ2, Igγ3, and Igγ4, respectively. Light chains are usually composed of polypeptide chains containing approximately 220 amino acids, and two types are known: the L type and the K type, called Igλ and Igκ, respectively. These two types of light chains can pair with any type of heavy chain. The peptide composition of the basic structure of antibody molecules consists of two identical heavy chains and two identical light chains, each connected by disulfide bonds and non-covalent bonds, and the molecular weight of antibody molecules is 150,000 to 190,000.

[0253] Antibody molecules have four intra-chain disulfide bonds in the heavy chain (five in Igμ and Igε) and two in the light chain, forming a loop every 100 to 110 amino acid residues. Their conformations between the loops are similar and are called structural units or domains. The domains located at the N-terminus of both the heavy chain and the light chain have different amino acid sequences even if the samples are from the same class (subclass) of the same animal species and are called variable regions. Each N-terminal domain of the heavy chain and the light chain is called "heavy chain variable region (VH) and light chain variable region (VL)", respectively. The amino acid sequence of the C-terminal domain downstream of the variable region is almost constant for each class or subclass and is called the "constant region". The heavy chain constant region (CH) is further divided into three domains from the N-terminal side: CH1, CH2, and CH3. The light chain constant region is called "CL".

[0254] In the Y-shaped structure of an antibody, the part corresponding to the vertical line in the lower half of the Y-shape is called the "Fc (fragment, crystallizable) region". The Fc region is composed of two "Fc polypeptides". The Fc polypeptide is composed of the CH2 and CH3 domains of the heavy chain. On the other hand, in the Y-shaped structure of an antibody, in the upper half of the Y-shaped structure, the part corresponding to the diagonal line forming the V-shaped part is called the "Fab (fragment, antigen-binding) region". The Fab region is composed of the VH and CH1 domains of the heavy chain and the light chain (VL and CL). The antibody binds to an antigen at the tip (antigen-binding site) of the V-shape formed by the two Fab regions. The Fab region and the Fc region of the heavy chain are connected by a region called the hinge region. In addition, the two heavy chains of the antibody are linked by disulfide bonds.

[0255] The "antigen-binding site" is composed of VH and VL, and the binding specificity to the antigen is determined by the amino acid sequence at this site. On the other hand, the biological activities (such as binding to complement and various Fc receptor-expressing cells) reflect the structural differences in the constant regions of each Ig class. The variability of the light and heavy chain variable regions has been understood to be limited to three small hypervariable regions, which exist in both chains and are called "complementary determining regions (CDRs)" and have relatively large changes in the amino acid sequence. On the other hand, the remaining part of the variable region is called the framework region (FR), which consists of FR1 to FR4 and has relatively small changes in the amino acid sequence.

[0256] There are three CDRs in each N-terminus of the heavy chain and the light chain, in the order of "CDR1", "CDR2", and "CDR3", which consist of about 5 to 10 amino acid residues. In particular, the CDR of the heavy chain is known to contribute to antigen binding. In addition, among CDR1 to CDR3, the contribution of CDR3 is known to be the largest.

[0257] The amino acid residue numbering of the antibodies used herein can be defined according to their numbering systems by specifying the Kabat numbering or the EU index (Kabat, Sequences of Proteins of Immunological Interest, 5th edition, 1991, NIH publication number 91-3242).

[0258] As used herein, an "antibody derivative" is a molecule that includes at least one polypeptide chain derived from a full-length immunoglobulin molecule and has antigen-binding activity. Representative antibody derivatives include single-chain variable region fragments (scFv), Fab fragments, Fab' fragments, and F(ab')2 fragments. An "scFv" is a monovalent antibody derivative composed of VH and VL linked by a linker. A "Fab fragment" is a monovalent antibody derivative composed of a light chain and a heavy chain fragment that includes VH, the CH1 domain, and a portion of the hinge region. An "F(ab')2Fab' fragment" is a monovalent antibody derivative composed of a light chain and a heavy chain fragment that includes VH, the CH1 domain, and a portion of the hinge region, and this portion of the hinge region includes cysteine residues that form the S-S bond between the heavy chains. The term "monovalent" means containing one antigen-binding site.

[0259] A "single-arm antibody" is also a type of antibody derivative. As used herein, a "single-arm antibody" is a monovalent antibody derivative that includes one Fab region and one Fc region, wherein the Fab region is linked to one of the two Fc polypeptides in the Fc region. The Fab region and one Fc polypeptide can be linked via a hinge region or a peptide linker. In one embodiment, the single-arm antibody includes one heavy chain (VH, CH1 domain, hinge region, Fc polypeptide (CH2 domain and CH3 domain)), one light chain (VL and CL), and an Fc polypeptide ( Figure 9 ). In one embodiment, the single-arm antibody is a single-arm antibody that includes one heavy chain, one light chain, and an Fc polypeptide, wherein a hinge region is added to the Fc polypeptide ( Figure 9 -left). In one embodiment, the single-arm antibody is a single-arm antibody composed of one heavy chain, one light chain, and an Fc polypeptide, wherein the CH1 domain is linked to the Fc polypeptide via a hinge region ( Figure 9 -middle). In one embodiment, the single-arm antibody is a single-arm antibody composed of one heavy chain, one light chain, and an Fc polypeptide ( Figure 9 –right). In one embodiment, the single-arm antibody can have two Fc polypeptides bound by an S-S bond between the heavy chains in the hinge region portion ( Figure 9 -left and middle), or these two Fc polypeptides can include modified residues or added cysteine residues in one or both of the Fc polypeptides to allow them to bind to each other ( Figure 9 -right).

[0260] The "anti-human NR1 antibody" in this article refers to an antibody that binds to human NR1. Whether it binds to human NR1 can be determined using well-known methods for measuring binding activity. Methods for measuring binding activity include, for example, enzyme-linked immunosorbent assay (ELISA). If ELISA is used, for example, a full-length or partial polypeptide of the human NR1 protein that can be produced based on the amino acid sequence shown in GenBank accession number NP_015566.1 is immobilized on an ELISA plate, and after reacting by adding a test antibody, it is reacted with a second antibody (such as an anti-IgG antibody labeled with an enzyme such as horseradish peroxidase (HRP)). After the reaction, the binding of the second antibody is identified by performing washing operations and measuring the activity using a reagent for detecting its activity, etc., and it can be confirmed whether the test antibody binds to human NR1. As a specific evaluation method, for example, the method described in Example 5 below can be used.

[0261] In addition to anti-human NR1, the anti-human NR1 antibody of the present invention also includes an antibody that binds to NR1 derived from another animal (for example, anti-monkey NR1), as long as the antibody binds to human NR1.

[0262] <The single-arm anti-human NR1 antibody of the present invention>

[0263] In this article, the "single-arm anti-human NR1 antibody" is a single-arm antibody that binds to human NR1.

[0264] The single-arm anti-human NR1 antibody of the present invention includes a single-arm anti-human NR1 antibody selected from any one of the following (1) to (5):

[0265] (1) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4;

[0266] (2) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, CDR2 consisting of the amino acid sequence from amino acid number 50 to 60 of SEQ ID NO: 6, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8;

[0267] (3) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12;

[0268] (4) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 16, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 16, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 16;

[0269] (5) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18, and the light - chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20.

[0270] In one embodiment, the single - arm anti - human NR1 antibody of the present invention is a single - arm anti - human NR1 antibody selected from any one of the following (6) to (10):

[0271] (6) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4;

[0272] (7) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0273] (8) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0274] (9) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0275] (10) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:18, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:20.

[0276] In one embodiment, the single - arm anti - human NR1 antibody of the present invention is a single - arm anti - human NR1 antibody selected from any one of the following (1) to (5):

[0277] (1) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, a CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and a CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and a CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4;

[0278] (2) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:6, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:6, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO:6; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:8, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:8, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO:8;

[0279] (3) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO:10; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:12;

[0280] (4) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:14, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:14, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:14; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:16, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:16, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:16;

[0281] (5) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20.

[0282] In one embodiment, the single - arm anti - human NR1 antibody of the present invention is a single - arm anti - human NR1 antibody selected from any one of the following (6) to (10):

[0283] (6) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region consists of the amino acid sequence shown in SEQ ID NO:2; the light chain comprises a light - chain variable region, and the light - chain variable region consists of the amino acid sequence shown in SEQ ID NO:4;

[0284] (7) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region consists of the amino acid sequence shown in SEQ ID NO:6; the light chain comprises a light - chain variable region, and the light - chain variable region consists of the amino acid sequence shown in SEQ ID NO:8;

[0285] (8) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:10, the light chain comprises a light - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:12;

[0286] (9) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:14, the light chain comprises a light - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:16;

[0287] (10) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:18, the light chain comprises a light - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:20.

[0288] As the heavy - chain constant region for deriving the CH1 domain and the Fc polypeptide of the single - arm anti - human NR1 antibody of the present invention, any one of the Igγ, Igμ, Igα, Igδ, or Igε constant regions can be selected. For example, Igγ can be selected from Igγ1, Igγ2, Igγ3, or Igγ4. In one embodiment, the single - arm anti - human NR1 antibody of the present invention comprises a CH1 domain and an Fc polypeptide derived from the Igγ1 constant region. In one embodiment, the single - arm anti - human NR1 antibody of the present invention comprises a CH1 domain and an Fc polypeptide derived from the human Igγ1 constant region.

[0289] As the CL of the single - arm anti - human NR1 antibody of the present invention, any one of the Igλ or Igκ constant regions can be selected. In one embodiment, the single - arm anti - human NR1 antibody of the present invention comprises a CL as the Igκ constant region. In one embodiment, the single - arm anti - human NR1 antibody of the present invention comprises a CL as the human Igκ constant region.

[0290] The heavy chain constant region of the single-chain anti-human NR1 antibody of the present invention may have mutations that reduce antibody-dependent cytotoxic activity or complement-dependent cytotoxic activity. L234A represents the substitution of leucine with alanine at amino acid position 234 according to the EU index in the human Igγ1 constant region. L235A represents the substitution of leucine with alanine at amino acid position 235 according to the EU index in the human Igγ1 constant region. The human Igγ1 constant region having the L234A and L235A amino acid mutations (hereinafter referred to as the "LALA mutation") includes, for example, the human Igγ1 constant region consisting of the amino acid sequence from amino acid number 121 to 450 in SEQ ID NO:22. It is known that the mutations reduce the antibody-dependent cytotoxic activity and complement-dependent cytotoxic activity of the antibody (Mol. Immunol., 1992, Vol. 29, pp. 633-639 / J. Immunol., 2000, Vol. 164, pp. 4178-4184). In addition, in murine antibodies, it is known that the D265A amino acid mutation in IgG1 reduces the antibody-dependent cytotoxic activity and complement-dependent cytotoxic activity of the antibody (J. Immunol., 2008, Vol. 181, pp. 6664-6669 / Nat. Med., 2000, Vol. 6, pp. 443-446). If one wants to observe the effect of an antibody with reduced antibody-dependent cytotoxic activity or complement-dependent cytotoxic activity in a murine model, a murine antibody with the D265A amino acid mutation can be used instead of the human antibody with the LALA mutation.

[0291] Other mutations based on known techniques can also be introduced into the heavy chain constant region of the single-chain anti-human NR1 antibody of the present invention. For example, the single-chain anti-human NR1 antibody of the present invention may have mutations based on the knobs-into-holes technology (hereinafter referred to as the "knobs-into-holes mutation"). The knobs-into-holes technology is a technique for effectively obtaining the desired heterodimeric antibody molecule by substituting the amino acid side chain present in the CH3 region of one heavy chain with a larger side chain (knob) and substituting the amino acid side chain present in the CH3 region of the other heavy chain with a smaller side chain (hole), such that the knob is located within the hole, thereby promoting the heterodimerization of the heavy chains (Nature, 1994, Vol. 372, pp. 379-383 / Nature Biotech., 1998, Vol. 16, pp. 677-681 / J. Mol. Biol., 1997, Vol. 270, pp. 26-35 / Proc. Natl. Acad. Sci. USA, 2013, Vol. 110, pp. E2987-E2996).

[0292] In one embodiment, the single-chain anti-human NR1 antibody of the present invention comprises an Fc region having the L234A and L235A amino acid mutations (LALA mutation).

[0293] In one embodiment, the single-chain anti-human NR1 antibody of the present invention comprises an Fc region having a knobs-into-holes mutation.

[0294] In one embodiment, the single-chain anti-human NR1 antibody of the present invention comprises an Fc region having L234A and L235A amino acid mutations (LALA mutation) and a knobs-into-holes mutation.

[0295] In one embodiment, the knobs-into-holes mutation of the single-chain anti-human NR1 antibody of the present invention is a T366W mutation in one Fc polypeptide forming its Fc region and T366S, L368A, and Y407V mutations in another Fc polypeptide forming its Fc region (see WO1998 / 050431).

[0296] As used herein, "human antibody" refers to an antibody having the amino acid sequence of a human immunoglobulin. As used herein, "humanized antibody" refers to an antibody in which some, most, or all of the amino acid residues other than the CDRs have been replaced with amino acid residues derived from a human immunoglobulin molecule. Humanized antibodies can be generated by reference to, for example, U.S. Patent No. 5,225,539 and U.S. Patent No. 6,180,370.

[0297] In one embodiment, the single-chain anti-human NR1 antibody of the present invention is a human antibody or a humanized antibody.

[0298] In one embodiment, the single-chain anti-human NR1 antibody of the present invention is a single-chain anti-human NR1 antibody comprising: a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22, a light chain consisting of the amino acid sequence shown in SEQ ID NO:24, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0299] The heavy chain comprising the amino acid sequence shown in SEQ ID NO:22 has an LALA mutation, and the mutation is located in the alanine residues at amino acid positions 237 and 238 in SEQ ID NO:22.

[0300] The heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22 has a knobs-into-holes mutation (T366S, L368A, and Y407V), and the mutation is located in the serine residue at amino acid position 369, the alanine residue at amino acid position 371, and the valine residue at amino acid position 410 in SEQ ID NO:22.

[0301] The Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42 has an LALA mutation, and the mutation is located in the alanine residues at amino acid positions 9 and 10 in SEQ ID NO: 42. The Fc polypeptide containing the amino acid sequence shown in SEQ ID NO: 42 also has a knobs-into-holes mutation (T366W), and the mutation is located in the tryptophan residue at amino acid position 141.

[0302] In one embodiment, the single-chain anti-human NR1 antibody of the present invention is a single-chain anti-human NR1 antibody comprising: a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0303] In one embodiment, the single-chain anti-human NR1 antibody of the present invention is a single-chain anti-human NR1 antibody comprising: a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0304] In one embodiment, the single-chain anti-human NR1 antibody of the present invention is a single-chain anti-human NR1 antibody comprising: a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0305] In one embodiment, the single-chain anti-human NR1 antibody of the present invention is a single-chain anti-human NR1 antibody comprising: a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0306] Like the heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, the heavy chains consisting of the amino acid sequences shown in SEQ ID NO: 26, 30, 34, and 38 also have LALA mutations and knobs-into-holes mutations (T366S, L368A, and Y407V).

[0307] In one embodiment, the single-arm anti-human NR1 antibody of the present invention competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4, and inhibits the NMDAR receptor endocytosis caused by pathogenic anti-human NR1 antibodies derived from patients with anti-NMDAR encephalitis.

[0308] As used herein, two antibodies "competitively" binding to the same antigen protein means that the epitopes on the antigen protein bound by each of the two antibodies are the same, or a part of the epitopes on the antigen protein bound by each of the two antibodies is common, or one of the antibodies binding to the epitope on the antigen protein changes the three-dimensional structure of the antigen protein, which contains the epitope portion bound by the other antibody on the antigen protein, resulting in the other antibody being no longer able to bind to the epitope portion on the antigen protein.

[0309] For example, those skilled in the art can obtain multiple single-arm anti-human NR1 antibodies, which are generated based on antibodies obtained by immunizing mice or rats with human NR1 polypeptides. Then, according to the methods disclosed in Examples 3-5, etc., herein, by screening with an anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4, and selecting an anti-human NR1 antibody that competitively binds to human NR1 with the anti-human NR1 antibody and inhibits the NMDAR receptor endocytosis caused by pathogenic anti-human NR1 antibodies, a single-arm anti-human NR1 antibody can be selected from the obtained various anti-human NR1 antibodies.

[0310] A "single-arm anti-human NR1 antibody that competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4" includes, for example, ABN_1-1-175, ABN_1-178.3, ABN_3-385, ABN_3-502 and their derivatives, which are identified as the results of the ELISA competitive assay described in Example 5 herein.

[0311] "A single - arm anti - human NR1 antibody that competitively binds to human NR1 with an anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4 and inhibits the NMDAR receptor endocytosis caused by pathogenic anti - human NR1 antibodies derived from patients with anti - NMDAR encephalitis" includes, for example, ABN_1 - 175, ABN_1 - 178, ABN_3 - 385, ABN_3 - 502, or a single - arm antibody comprising the antigen - binding site of P003 - 102 described in Example 1, or more specifically, the single - arm anti - human NR1 antibodies ABN - 1 to - 5 produced in Example 2 or Example 6 herein.

[0312] When antibodies are expressed in cells, it is known that they undergo post - translational modifications. Examples of post - translational modifications include cleavage of the heavy - chain C - terminal lysine by carboxypeptidase, modification of the heavy - chain and light - chain N - terminal glutamine or glutamate with pyroglutamate by pyroglutamylation, glycosylation, oxidation, deamidation, and glycation, etc. Such post - translational modifications are known to occur in various antibodies (J. Pharm. Sci., 2008, Vol. 97, pp. 2426 - 2447).

[0313] Examples of the single - arm anti - human NR1 antibodies of the present invention produced by post - translational modifications include single - arm anti - human NR1 antibodies that have undergone N - terminal pyroglutamylation of the heavy - chain variable region and / or deletion of the heavy - chain C - terminal lysine. Such post - translational modifications by N - terminal pyroglutamylation or C - terminal lysine deletion are known in the art not to affect the activity of the antibody (Anal. Biochem., 2006, Vol. 348, pp. 24 - 39).

[0314] In one embodiment, the single - arm anti - human NR1 antibody is produced by post - translational modification.

[0315] In one embodiment, the single - arm anti - human NR1 antibody is produced by post - translational modifications of N - terminal pyroglutamylation of the heavy - chain variable region of the single - arm anti - human NR1 antibody of the present invention and / or deletion of the heavy - chain C - terminal lysine.

[0316] Based on the VH and VL sequence information of the single - arm anti - human NR1 antibody of the present invention disclosed herein, etc., the single - arm anti - human NR1 antibody of the present invention can be easily produced by those skilled in the art using methods well - known in the art. The single - arm anti - human NR1 antibody of the present invention can be produced by, for example, the methods described in the following "Method for producing the single - arm anti - human NR1 antibody of the present invention, and the single - arm anti - human NR1 antibody thus produced", but is not limited thereto.

[0317] If needed, the single - arm anti - human NR1 antibody of the present invention, after further purification, is formulated by standard methods and can be used for the treatment of anti - NMDAR encephalitis, etc.

[0318] <The polynucleotide of the present invention>

[0319] The polynucleotide of the present invention includes a polynucleotide comprising a nucleotide sequence encoding the heavy chain variable region of the single-arm anti-human NR1 antibody of the present invention and a polynucleotide comprising a nucleotide sequence encoding the light chain variable region of the single-arm anti-human NR1 antibody of the present invention.

[0320] In one embodiment, the polynucleotide of the present invention is a polynucleotide comprising the sequences described in the following (a) and / or (b):

[0321] (a) Nucleotide sequences encoding the heavy chain variable region of the single-arm anti-human NR1 antibody selected from the following (a-1) to (a-4):

[0322] (a-1) A heavy chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 6, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6;

[0323] (a-2) A heavy chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10;

[0324] (a-3) A heavy chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14;

[0325] (a-4) A heavy chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 18; and / or

[0326] (b) Nucleotide sequences encoding the light chain variable region of the single-arm anti-human NR1 antibody selected from the following (b-1) to (b-4):

[0327] (b-1) A light chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:8, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:8, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO:8;

[0328] (b-2) A light chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:12;

[0329] (b-3) A light chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:16, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:16, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:16;

[0330] (b-4) A light chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO:20.

[0331] In one embodiment, the polynucleotide of the present invention is a polynucleotide comprising the nucleotide sequences described in the following (a) and / or (b):

[0332] (a) Nucleotide sequences encoding the heavy chain variable region of a single-arm anti-human NR1 antibody selected from the following (a-1) to (a-4):

[0333] (a-1) A heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6;

[0334] (a-2) A heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10;

[0335] (a-3) A heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14;

[0336] (a-4) A heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:18; and / or

[0337] (b) The nucleotide sequence encoding the light chain variable region of the encoded single-chain anti-human NR1 antibody selected from the following (b-1) to (b-4):

[0338] (b-1) The light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0339] (b-2) The light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0340] (b-3) The light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0341] (b-4) The light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:20.

[0342] In one embodiment, the polynucleotide of the present invention is a polynucleotide comprising the nucleotide sequence described in the following (a) and / or (b):

[0343] (a) The nucleotide sequence encoding the heavy chain of the encoded single-chain anti-human NR1 antibody selected from the following (a-1) to (a-4):

[0344] (a-1) A heavy chain comprising a heavy chain variable region, the heavy chain variable region comprising CDR1 consisting of the amino acid sequence of amino acid numbers 31 to 35 of SEQ ID NO:6, CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 66 of SEQ ID NO:6, and CDR3 consisting of the amino acid sequence of amino acid numbers 99 to 110 of SEQ ID NO:6;

[0345] (a-2) A heavy chain comprising a heavy chain variable region, the heavy chain variable region comprising CDR1 consisting of the amino acid sequence of amino acid numbers 31 to 35 of SEQ ID NO:10, CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 66 of SEQ ID NO:10, and CDR3 consisting of the amino acid sequence of amino acid numbers 99 to 111 of SEQ ID NO:10;

[0346] (a-3) A heavy chain comprising a heavy chain variable region, the heavy chain variable region comprising CDR1 consisting of the amino acid sequence of amino acid numbers 31 to 36 of SEQ ID NO:14, CDR2 consisting of the amino acid sequence of amino acid numbers 51 to 66 of SEQ ID NO:14, and CDR3 consisting of the amino acid sequence of amino acid numbers 99 to 108 of SEQ ID NO:14;

[0347] (a - 4) A heavy chain comprising a heavy chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence of amino acid numbers 31 to 36 of SEQ ID NO: 18, a CDR2 consisting of the amino acid sequence of amino acid numbers 51 to 66 of SEQ ID NO: 18, and a CDR3 consisting of the amino acid sequence of amino acid numbers 99 to 108 of SEQ ID NO: 18; and / or

[0348] (b) A nucleotide sequence encoding a light chain of a single - arm anti - human NR1 antibody selected from the following (b - 1) to (b - 4):

[0349] (b - 1) A light chain comprising a light chain variable region, wherein the light chain variable region comprises a CDR1 consisting of the amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 8, a CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence of amino acid numbers 89 to 98 of SEQ ID NO: 8;

[0350] (b - 2) A light chain comprising a light chain variable region, wherein the light chain variable region comprises a CDR1 consisting of the amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 12, a CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 12, and a CDR3 consisting of the amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO: 12;

[0351] (b - 3) A light chain comprising a light chain variable region, wherein the light chain variable region comprises a CDR1 consisting of the amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 16, and a CDR3 consisting of the amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO: 16;

[0352] (b - 4) A light chain comprising a light chain variable region, wherein the light chain variable region comprises a CDR1 consisting of the amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 20, a CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 20, and a CDR3 consisting of the amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO: 20.

[0353] In one embodiment, the polynucleotide of the present invention comprises the nucleotide sequences described in the following (a) and / or (b):

[0354] (a) A nucleotide sequence encoding a heavy chain variable region of a single - arm anti - human NR1 antibody selected from the following (a - 1) to (a - 4):

[0355] (a-1) A heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6;

[0356] (a-2) A heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10;

[0357] (a-3) A heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14;

[0358] (a-4) A heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:18; and / or

[0359] (b) A nucleotide sequence encoding a light chain variable region of a single-arm anti-human NR1 antibody selected from the following (b-1) to (b-4):

[0360] (b-1) A light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0361] (b-2) A light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0362] (b-3) A light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0363] (b-4) A light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:20.

[0364] In one embodiment, the polynucleotide of the present invention comprises the nucleotide sequences described in the following (a) and / or (b):

[0365] (a) A nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26; and / or

[0366] (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28.

[0367] The polynucleotide encoding the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 includes, for example, a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:25.

[0368] The polynucleotide encoding the light chain consisting of the amino acid sequence shown in SEQ ID NO:28 includes, for example, a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:27.

[0369] In one embodiment, the polynucleotide of the present invention comprises the nucleotide sequence described in the following (a) and / or (b):

[0370] (a) A nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30; and / or

[0371] (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32.

[0372] Polynucleotides comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:30 include, for example, polynucleotides comprising the nucleotide sequence shown in SEQ ID NO:29.

[0373] Polynucleotides comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:32 include, for example, polynucleotides comprising the nucleotide sequence shown in SEQ ID NO:31.

[0374] In one embodiment, the polynucleotide of the present invention comprises the nucleotide sequence described in the following (a) and / or (b):

[0375] (a) A nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:34; and / or

[0376] (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:36.

[0377] Polynucleotides comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:34 include, for example, polynucleotides comprising the nucleotide sequence shown in SEQ ID NO:33.

[0378] Polynucleotides comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:36 include, for example, polynucleotides comprising the nucleotide sequence shown in SEQ ID NO:35.

[0379] In one embodiment, the polynucleotide of the present invention comprises the nucleotide sequence described in the following (a) and / or (b):

[0380] (a) A nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:38; and / or

[0381] (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:40.

[0382] A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:38 includes, for example, a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:37.

[0383] A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:40 includes, for example, a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:39.

[0384] In one embodiment, the polynucleotide of the present invention may further comprise a nucleotide sequence encoding an Fc polypeptide, and in one embodiment, the polynucleotide of the present invention may comprise a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0385] In one embodiment, the polynucleotide of the present invention comprises the nucleotide sequences described in the following (a) and / or (b) and (c):

[0386] (a) A nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22;

[0387] (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24.

[0388] (c) A nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42.

[0389] A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:22 includes, for example, a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:21.

[0390] A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:24 includes, for example, a polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:23.

[0391] In one embodiment, the polynucleotide of the present invention is a polynucleotide comprising a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody and / or a nucleotide sequence encoding a light chain of a single-arm anti-human NR1 antibody, wherein the antibody competes with an anti-human NR1 antibody comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4 for binding to human NR1, and inhibits NMDAR endocytosis induced by pathogenic anti-human NR1 antibodies derived from patients with anti-NMDAR encephalitis.

[0392] The polynucleotides of the present invention can be readily produced by those skilled in the art based on their nucleotide sequences and by using methods well known in the art. For example, the polynucleotides of the present invention can be synthesized by utilizing gene synthesis methods known in the art. Such gene synthesis methods that can be used include various methods known to those skilled in the art, such as the method for synthesizing antibody genes described in WO90 / 07861.

[0393] <The expression vectors of the present invention>

[0394] The expression vectors of the present invention include expression vectors comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-chain anti-human antibody, expression vectors comprising a polynucleotide containing a nucleotide sequence encoding a light chain variable region of a single-chain anti-human antibody, and expression vectors comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-chain anti-human antibody of the present invention and a polynucleotide containing a nucleotide sequence encoding a light chain variable region of said antibody. In addition, the expression vectors of the present invention include expression vectors comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-chain anti-human antibody of the present invention, expression vectors comprising a polynucleotide containing a nucleotide sequence encoding a light chain of a single-chain anti-human antibody of the present invention, and expression vectors comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-chain anti-human antibody of the present invention and a polynucleotide containing a nucleotide sequence encoding a light chain of said antibody. The expression vectors of the present invention may comprise a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide.

[0395] In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, a CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and a CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2. And the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and a CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO:2, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO:4. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, a CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and a CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2. And the light chain comprises a light chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and a CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2, and the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain consists of the amino acid sequence shown in SEQ ID NO:22, and the light chain consists of the amino acid sequence shown in SEQ ID NO:24.

[0396] In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 6, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region, wherein the heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO: 6, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 8. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain, wherein the heavy chain comprises a heavy chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 6, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6, and the light chain comprises a light chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain, wherein the heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 6, and the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 8. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 26, and the light chain consists of the amino acid sequence shown in SEQ ID NO: 28.

[0397] In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10. And the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO: 10, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 12. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10. And the light chain comprises a light chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10, and the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 12.In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 30, and the light chain consists of the amino acid sequence shown in SEQ ID NO: 32.

[0398] In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14. And the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 16, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 16. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO: 14, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 16. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14. And the light chain comprises a light chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 16, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 16. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 14, and the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 16.In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 34, and the light chain consists of the amino acid sequence shown in SEQ ID NO: 36.

[0399] In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 18. And the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 20. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region and / or a polynucleotide containing a nucleotide sequence encoding a light chain variable region. The heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO: 18, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 20. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 18. And the light chain comprises a light chain variable region containing a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 20. In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain. The heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 18, and the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 20.In one embodiment, the expression vector of the present invention comprises a polynucleotide containing a nucleotide sequence encoding a heavy chain and / or a polynucleotide containing a nucleotide sequence encoding a light chain, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 38, and the light chain consists of the amino acid sequence shown in SEQ ID NO: 40.

[0400] In one embodiment, the expression vector of the present invention may further comprise a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide. In one embodiment, the expression vector of the present invention may further comprise a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide shown in SEQ ID NO: 42.

[0401] In one embodiment, the expression vector of the present invention is an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody and / or a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody, wherein the antibody competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 4, and inhibits NMDAR endocytosis induced by pathogenic anti-human NR1 antibodies derived from patients with anti-NMDAR encephalitis.

[0402] In one embodiment, the expression vector of the present invention is an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody and / or a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody and the Fc polypeptide of the antibody, wherein the antibody competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 4, and inhibits NMDAR endocytosis induced by pathogenic anti-human NR1 antibodies derived from patients with anti-NMDAR encephalitis.

[0403] Expression vectors for expressing the polynucleotides of the present invention include, but are not limited to, expression vectors that can express polynucleotides containing nucleotide sequences encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention, polynucleotides containing nucleotide sequences encoding the light chain of the single-arm anti-human NR1 antibody of the present invention, or polynucleotides containing nucleotide sequences encoding an Fc polypeptide that binds to the heavy chain to form an Fc region, and produce the polypeptides encoded by them in various host cells such as eukaryotic cells (e.g., animal cells, insect cells, plant cells, and yeast) and / or prokaryotic cells (e.g., Escherichia coli). Such expression vectors include, for example, plasmid vectors, viral vectors (e.g., adenovirus and retrovirus), etc. In one embodiment, pEE6.4 or pEE12.4 (Lonza) can be used as the expression vector.

[0404] The expression vectors of the present invention may contain a promoter operably coupled to the polynucleotides of the present invention. Promoters for expressing the polynucleotides of the present invention in animal cells include, for example, promoters of viral origin (such as CMV, RSV, and SV40), actin promoter, EF (elongation factor) 1α promoter, heat shock promoter, etc. Promoters for expressing the polynucleotides of the present invention in bacteria (e.g., Escherichia coli) include, for example, trp promoter, lac promoter, λPL promoter, tac promoter, etc. Promoters for expressing the polynucleotides of the present invention in yeast include, for example, GAL 1 promoter, GAL 10 promoter, PH05 promoter, PGK promoter, GAP promoter, ADH promoter, etc.

[0405] When using animal cells, insect cells, or yeast as host cells, the expression vectors of the present invention may include a start codon, a stop codon, an enhancer sequence, 5' and 3' untranslated regions of the gene encoding the single-arm anti-human NR1 antibody or its heavy or light chain of the present invention, a secretion signal sequence, a splice junction, a polyadenylation site, or a replicable unit, etc. When using Escherichia coli as the host cell, the expression vectors of the present invention may include a start codon, a stop codon, a terminator region, and a replicable unit. According to the purpose, the expression vectors of the present invention may include commonly used selectable markers (e.g., tetracycline resistance gene, ampicillin resistance gene, kanamycin resistance gene, neomycin resistance gene, and dihydrofolate reductase gene).

[0406] <The transformed host cells of the present invention>

[0407] The transformed host cells of the present invention include transformed host cells having the expression vectors of the present invention. The transformed host cells of the present invention include host cells selected from the following (1)-(4):

[0408] (1) A transformed host cell having an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain variable region of the single-arm anti-human NR1 antibody of the present invention;

[0409] (2) A transformed host cell having an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain variable region of the single-arm anti-human NR1 antibody of the present invention;

[0410] (3) A transformed host cell having an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain variable region of the single-arm anti-human NR1 antibody of the present invention, and a polynucleotide containing a nucleotide sequence encoding the light chain variable region of said antibody;

[0411] (4) A transformed host cell having an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain variable region of the single-arm anti-human NR1 antibody of the present invention, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain variable region of said antibody.

[0412] The transformed host cells of the present invention further include host cells selected from the following (a)-(d):

[0413] (a) A transformed host cell having an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention;

[0414] (b) A transformed host cell having an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the single-arm anti-human NR1 antibody of the present invention;

[0415] (c) A transformed host cell having an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention, and a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody;

[0416] (d) A transformed host cell having an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody.

[0417] In one embodiment, the expression vector in the host cells included in the above (a)-(d) may further comprise a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide. In one embodiment, the host cells in the above (a)-(d) may further comprise an expression vector, said expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide.

[0418] Examples of the transformed host cells of the present invention include transformed host cells having an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-chain anti-human NR1 antibody of the present invention, a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of the antibody; transformed host cells having an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-chain anti-human NR1 antibody of the present invention, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of the antibody; and transformed host cells having an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-chain anti-human NR1 antibody of the present invention and a polynucleotide containing a nucleotide sequence encoding a light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of the antibody.

[0419] In one embodiment, the host cell of the present invention is a transformed host cell having an expression vector selected from the following (a)-(d):

[0420] (a) an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-chain anti-human NR1 antibody of the present invention listed in any one of the following (1) to (8);

[0421] (b) an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain variable region of a single-chain anti-human NR1 antibody of the present invention listed in any one of the following (1) to (8); (c) an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-chain anti-human NR1 antibody of the present invention listed in any one of the following (1) to (8) and a polynucleotide containing a nucleotide sequence encoding a light chain variable region of the antibody;

[0422] (d) an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-chain anti-human NR1 antibody of the present invention listed in any one of the following (1) to (8), and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain variable region of the antibody:

[0423] (1) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, CDR2 consisting of the amino acid sequence from amino acid number 50 to 60 of SEQ ID NO: 6, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8;

[0424] (2) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12;

[0425] (3) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 16, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 16, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 16;

[0426] (4) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20;

[0427] (5) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0428] (6) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0429] (7) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0430] (8) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:18, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:20.

[0431] In one embodiment, the host cell of the present invention is a transformed host cell having an expression vector selected from:

[0432] (a) An expression vector comprising a polynucleotide encoding a heavy chain of the single-arm anti-human NR1 antibody of the present invention listed in any one of (1) to (8) below;

[0433] (b) An expression vector comprising a polynucleotide encoding a light chain of the single-arm anti-human NR1 antibody of the present invention listed in any one of (1) to (8) below;

[0434] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody of the present invention listed in any one of the following (1) to (8) and a polynucleotide containing a nucleotide sequence encoding the light chain variable region of said antibody;

[0435] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of a single-arm anti-human NR1 antibody of the present invention listed in any one of the following (1) to (8) and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of said antibody:

[0436] (1) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, the heavy chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 6, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6; the light chain comprises a light chain variable region, the light chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8;

[0437] (2) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, the heavy chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10; the light chain comprises a light chain variable region, the light chain variable region comprising a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12;

[0438] (3) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 16, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 16;

[0439] (4) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 18; the light chain comprises a light chain variable region, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 20;

[0440] (5) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO: 6, and the light chain comprises a light chain variable region, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 8;

[0441] (6) An anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region consists of the amino acid sequence shown in SEQ ID NO: 10, and the light chain comprises a light chain variable region, and the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 12;

[0442] (7) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:14, the light chain comprises a light - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:16;

[0443] (8) An anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:18, and the light chain comprises a light - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:20.

[0444] In one embodiment, the host cell of the present invention is a transformed host cell having an expression vector selected from the following (a) - (i):

[0445] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single - arm anti - human NR1 antibody of the present invention listed in any one of (1) to (10) below and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0446] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the single - arm anti - human NR1 antibody of the present invention listed in any one of (1) to (10) below and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0447] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single - arm anti - human NR1 antibody of the present invention listed in any one of (1) to (10) below and a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0448] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single - arm anti - human NR1 antibody of the present invention listed in any one of (1) to (10) below, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0449] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single - arm anti - human NR1 antibody of the present invention listed in any one of (1) to (10) below and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0450] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the single-arm anti-human NR1 antibody of the present invention listed in any one of the following (1) to (10), and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0451] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention listed in any one of the following (1) to (10) and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0452] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention listed in any one of the following (1) to (10), and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0453] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-arm anti-human NR1 antibody of the present invention listed in any one of the following (1) to (10), a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody:

[0454] (1) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2; the light chain comprises a light chain variable region, and the light chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4;

[0455] (2) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises CDR1 consisting of the amino - acid sequence from amino - acid number 31 to 35 of SEQ ID NO:6, CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 66 of SEQ ID NO:6, and CDR3 consisting of the amino - acid sequence from amino - acid number 99 to 110 of SEQ ID NO:6; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino - acid sequence from amino - acid number 24 to 34 of SEQ ID NO:8, CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 56 of SEQ ID NO:8, and CDR3 consisting of the amino - acid sequence from amino - acid number 89 to 98 of SEQ ID NO:8;

[0456] (3) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises CDR1 consisting of the amino - acid sequence from amino - acid number 31 to 35 of SEQ ID NO:10, CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 66 of SEQ ID NO:10, and CDR3 consisting of the amino - acid sequence from amino - acid number 99 to 111 of SEQ ID NO:10; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino - acid sequence from amino - acid number 24 to 34 of SEQ ID NO:12, CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 56 of SEQ ID NO:12, and CDR3 consisting of the amino - acid sequence from amino - acid number 89 to 97 of SEQ ID NO:12;

[0457] (4) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises CDR1 consisting of the amino - acid sequence from amino - acid number 31 to 36 of SEQ ID NO:14, CDR2 consisting of the amino - acid sequence from amino - acid number 51 to 66 of SEQ ID NO:14, and CDR3 consisting of the amino - acid sequence from amino - acid number 99 to 108 of SEQ ID NO:14; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino - acid sequence from amino - acid number 24 to 34 of SEQ ID NO:16, CDR2 consisting of the amino - acid sequence from amino - acid number 50 to 56 of SEQ ID NO:16, and CDR3 consisting of the amino - acid sequence from amino - acid number 89 to 97 of SEQ ID NO:16;

[0458] (5) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18; the light chain comprises a light chain variable region, and the light chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20;

[0459] (6) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region is composed of the amino acid sequence shown in SEQ ID NO:2, and the light chain comprises a light chain variable region, and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO:4;

[0460] (7) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region is composed of the amino acid sequence shown in SEQ ID NO:6, and the light chain comprises a light chain variable region, and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO:8;

[0461] (8) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region is composed of the amino acid sequence shown in SEQ ID NO:10, and the light chain comprises a light chain variable region, and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO:12;

[0462] (9) A single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy chain variable region, and the heavy chain variable region is composed of the amino acid sequence shown in SEQ ID NO:14, and the light chain comprises a light chain variable region, and the light chain variable region is composed of the amino acid sequence shown in SEQ ID NO:16;

[0463] (10) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:18, the light chain comprises a light - chain variable region consisting of the amino - acid sequence shown in SEQ ID NO:20.

[0464] In one embodiment, the host cell of the present invention is a transformed host cell having an expression vector selected from the following (a) - (i):

[0465] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino - acid sequence shown in SEQ ID NO:22, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino - acid sequence shown in SEQ ID NO:42;

[0466] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino - acid sequence shown in SEQ ID NO:22 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino - acid sequence shown in SEQ ID NO:42;

[0467] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino - acid sequence shown in SEQ ID NO:24, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino - acid sequence shown in SEQ ID NO:42;

[0468] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino - acid sequence shown in SEQ ID NO:24 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino - acid sequence shown in SEQ ID NO:42;

[0469] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino - acid sequence shown in SEQ ID NO:22 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino - acid sequence shown in SEQ ID NO:24, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino - acid sequence shown in SEQ ID NO:42;

[0470] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 24;

[0471] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 24 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0472] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 24, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0473] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 24, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0474] In one embodiment, the host cell of the present invention is a transformed host cell having one or more expression vectors selected from the following:

[0475] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26;

[0476] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28;

[0477] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28;

[0478] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28;

[0479] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0480] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0481] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0482] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0483] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:26 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:28, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:42;

[0484] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28;

[0485] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0486] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0487] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0488] In one embodiment, the host cell of the present invention is a transformed host cell having one or more expression vectors selected from the following:

[0489] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30;

[0490] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0491] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0492] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0493] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0494] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0495] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0496] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0497] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0498] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32;

[0499] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0500] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an expression vector comprising a polynucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0501] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0502] In one embodiment, the host cell of the present invention is a transformed host cell having one or more expression vectors selected from the following:

[0503] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34;

[0504] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0505] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0506] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0507] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0508] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0509] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0510] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0511] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0512] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36;

[0513] (k) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, and an expression vector comprising a polynucleotide having a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36 and a polynucleotide having a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0514] (l) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, an expression vector comprising a polynucleotide having a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an expression vector comprising a polynucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0515] (m) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, a polynucleotide having a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and a polynucleotide having a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0516] In one embodiment, the host cell of the present invention is a transformed host cell having one or more expression vectors selected from the following:

[0517] (a) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38;

[0518] (b) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0519] (c) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide having a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0520] (d) An expression vector comprising a polynucleotide having a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and an expression vector comprising a polynucleotide having a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0521] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0522] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0523] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0524] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0525] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0526] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40;

[0527] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0528] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an expression vector comprising a polynucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42;

[0529] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42.

[0530] In one embodiment, the host cell of the present invention is a transformed host cell having an expression vector encoding the heavy chain and / or light chain of a single-arm anti-human NR1 antibody, the single-arm anti-human NR1 antibody competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 4, and inhibits NMDAR endocytosis caused by pathogenic anti-human NR1 antibodies derived from anti-NMDAR encephalitis patients, wherein the host cell of the present invention is a transformed host cell having one or more expression vectors selected from the following:

[0531] (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody;

[0532] (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0533] (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0534] (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0535] (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0536] (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0537] (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0538] (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0539] (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0540] (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0541] (k) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0542] (l) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0543] (m) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody, a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody.

[0544] There are no particular limitations on the transformed host cells, as long as they are compatible with the expression vectors used, can be transformed with one or more expression vectors, and express the antibody. Transformed host cells include, for example, various cells such as natural cells or artificially established cells commonly used in the technical field of the present invention (such as animal cells (such as CHOK1SV1 cells), insect cells (such as Sf9), prokaryotic cells (such as Escherichia coli, etc.), yeasts (such as Saccharomyces, Pichia, etc.), etc.). In one embodiment, cultured cells such as CHOK1SV cells, CHO-DG44 cells, HEK293 cells, and NS0 cells, etc. can be used as host cells.

[0545] Methods for transforming host cells include the calcium phosphate method, electroporation method, etc.

[0546] <Method for producing the single-chain anti-human NR1 antibody of the present invention, and the single-chain anti-human NR1 antibody thus produced>

[0547] Methods for producing the single-chain anti-human NR1 antibody include methods of culturing the transformed host cells of the present invention, expressing the single-chain anti-human NR1 antibody, and producing the single-chain anti-human NR1 antibody. In one embodiment, the method for producing the single-chain anti-human NR1 antibody of the present invention includes culturing host cells selected from the following (a)-(e) and expressing the single-chain anti-human NR1 antibody:

[0548] (a) A host cell transformed with an expression vector, the expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody of the present invention, a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0549] (b) A host cell transformed with an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody and a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody;

[0550] (c) A host cell transformed with an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody;

[0551] (d) A host cell transformed with an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the single-chain anti-human NR1 antibody and a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the antibody;

[0552] (e) A host cell transformed with an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody of the present invention, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the light chain of the antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding the Fc polypeptide of the antibody.

[0553] The method for producing the single-chain anti-human NR1 antibody of the present invention is not particularly limited as long as the method includes culturing the transformed host cell of the present invention and expressing the single-chain anti-human NR1 antibody.

[0554] The transformed host cells can be cultured by known methods. The culture conditions are appropriately selected, for example, temperature, pH of the medium, and culture time. If the host cells are animal cells, media such as MEM medium (Science, 1959, Vol. 130, pp. 432-437) containing about 5-20% fetal bovine serum, DMEM medium (Virology, 1959, Vol. 8, p. 396), RPMI 1640 medium (J. Am. Med. Assoc., 1967, Vol. 199, p. 519), 199 medium (Exp. Biol. Med., 1950, Vol. 73, pp. 1-8), etc. can be used as the medium. The pH of the medium is, for example, about 6-8, and the culture is carried out at about 30-40 °C for about 15-336 hours periodically, with ventilation and stirring if necessary. If the host cells are insect cells, media such as Grace medium (Proc. Natl. Acad. Sci. USA., 1985, Vol. 82, p. 8404) containing fetal bovine serum can be used as the medium, for example. The pH of the medium is, for example, about 5-8, and the culture is carried out at about 20-40 °C for about 15-100 hours periodically, with ventilation and stirring if necessary. If the host cells are Escherichia coli or yeast, a liquid medium containing, for example, a nutrient source is suitable as the medium. The nutrient medium contains, for example, a carbon source, an inorganic or organic nitrogen source required for the growth of the transformed host cells. Carbon sources include, for example, glucose, dextran, soluble starch, and sucrose, etc., while inorganic or organic nitrogen sources include, for example, ammonium salts, nitrates, amino acids, corn steep liquor, peptone, casein, meat extract, soybean powder, and potato extract, etc. If necessary, it can contain other nutrients (for example, inorganic salts (for example, calcium chloride, sodium dihydrogen phosphate, magnesium chloride) and vitamins), antibiotics (for example, tetracycline, neomycin, ampicillin, kanamycin), etc. The pH of the medium is, for example, about 5-8. If the host cells are Escherichia coli, media such as LB medium and M9 medium (Molecular Cloning, Cold Spring Harbor Laboratory, Vol. 3, A2.2) can be used as the medium, for example. The culture is carried out at about 14-43 °C for about 3-24 hours periodically, with ventilation and stirring if necessary. If the host cells are yeast, media such as Burkholder minimal medium (Proc. Natl. Acad. Sci. USA., 1980, Vol. 77, p. 4505) can be used as the medium, for example. The culture is carried out at about 20-35 °C for about 14-144 hours periodically, with ventilation and stirring if necessary. The single-arm anti-human NR1 antibody of the present invention can be expressed by the above culture.

[0555] In addition to culturing transformed host cells to express the single-chain anti-human NR1 antibody of the present invention, the method for producing the single-chain anti-human NR1 antibody of the present invention may further include recovering, isolating, or purifying the single-chain anti-human NR1 antibody from the transformed host cells. Methods for isolation or purification include, for example, methods utilizing solubility such as salting out and solvent precipitation; methods utilizing molecular weight differences such as dialysis, ultrafiltration, and gel filtration; methods utilizing charge such as ion exchange chromatography and hydroxyapatite chromatography; methods utilizing specific affinity such as affinity chromatography; methods utilizing hydrophobicity differences such as reverse-phase high performance liquid chromatography; and methods utilizing isoelectric point differences such as isoelectric focusing electrophoresis, etc. In one embodiment, the antibody accumulated in the culture supernatant can be purified by various chromatography methods, such as column chromatography using a protein A column or a protein G column.

[0556] In the method for producing the single-chain anti-human NR1 antibody of the present invention, the single-chain anti-human NR1 antibody can be effectively obtained by introducing knobs-into-holes mutations into the CH3 of the heavy chain of the single-chain anti-human NR1 antibody and the CH3 of the Fc polypeptide that combines with the heavy chain to form the Fc region, respectively.

[0557] The single-chain anti-human NR1 antibody of the present invention also includes the single-chain anti-human NR1 antibody produced by the method for producing the single-chain anti-human NR1 antibody of the present invention.

[0558] <The pharmaceutical composition, etc. of the present invention>

[0559] The pharmaceutical composition of the present invention comprises a composition containing the single-chain anti-human NR1 antibody of the present invention and a pharmaceutically acceptable excipient. The pharmaceutical composition of the present invention can be prepared by conventional methods using excipients commonly used in the art, namely pharmaceutical excipients or drug carriers, etc. Dosage forms of these pharmaceutical compositions include, for example, parenteral dosage forms such as injection, infusion, etc., and the pharmaceutical composition can be administered by intravenous administration, subcutaneous administration, etc. When formulating the pharmaceutical composition, excipients, carriers, additives, etc. for these dosage forms can be used within a pharmaceutically acceptable range.

[0560] The pharmaceutical composition of the present invention may contain multiple single-chain anti-human NR1 antibodies of the present invention. For example, the present invention includes a single-chain anti-human NR1 antibody without C-terminal lysine deletion and N-terminal pyroglutamylation, and / or a single-chain anti-human NR1 antibody with C-terminal lysine deletion and N-terminal pyroglutamylation.

[0561] In one embodiment, the pharmaceutical composition of the present invention comprises a single-chain anti-human NR1 antibody selected from any one of the following (1) to (5) and / or a single-chain anti-human NR1 antibody produced by post-translational modification of the single-chain anti-human NR1 antibody:

[0562] (1) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO:2, CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO:2, and CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO:2, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO:4, CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO:4, and CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO:4;

[0563] (2) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:6, CDR2 consisting of the amino acid sequence from amino acid number 50 to 60 of SEQ ID NO:6, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO:6, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:8, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:8, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO:8;

[0564] (3) A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:10, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:10, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO:10, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:12, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:12, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:12;

[0565] (4) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:14, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:14, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:14, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:16, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:16, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:16;

[0566] (5) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20.

[0567] In one embodiment, the pharmaceutical composition of the present invention comprises a single - arm anti - human NR1 antibody selected from any one of the following (1) to (5) and / or a single - arm anti - human NR1 antibody produced by post - translational modification of the single - arm anti - human NR1 antibody:

[0568] (1) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4;

[0569] (2) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6 and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0570] (3) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10 and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0571] (4) A single-chain anti-human NR1 antibody, which comprises a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 14, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 16;

[0572] (5) A single-chain anti-human NR1 antibody, which comprises a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 18, and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 20.

[0573] In one embodiment, the pharmaceutical composition of the present invention comprises a single-chain anti-human NR1 antibody selected from any one of the following (1) to (5) and / or a single-chain anti-human NR1 antibody produced by post-translational modification of the single-chain anti-human NR1 antibody:

[0574] (1) A single-chain anti-human NR1 antibody comprising a heavy chain, a light chain and an Fc polypeptide, the heavy chain comprising a heavy-chain variable region, the heavy-chain variable region comprising CDR1 consisting of the amino acid sequence from amino acid number 26 to 34 of SEQ ID NO: 2, CDR2 consisting of the amino acid sequence from amino acid number 52 to 58 of SEQ ID NO: 2, and CDR3 consisting of the amino acid sequence from amino acid number 97 to 109 of SEQ ID NO: 2, the light chain comprising a light-chain variable region, the light-chain variable region comprising CDR1 consisting of the amino acid sequence from amino acid number 26 to 33 of SEQ ID NO: 4, CDR2 consisting of the amino acid sequence from amino acid number 51 to 53 of SEQ ID NO: 4, and CDR3 consisting of the amino acid sequence from amino acid number 92 to 100 of SEQ ID NO: 4;

[0575] (2) A single-chain anti-human NR1 antibody comprising a heavy chain, a light chain and an Fc polypeptide, the heavy chain comprising a heavy-chain variable region, the heavy-chain variable region comprising CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 6, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 6, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 110 of SEQ ID NO: 6, the light chain comprising a light-chain variable region, the light-chain variable region comprising CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 8, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 8, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 98 of SEQ ID NO: 8;

[0576] (3) A single - arm anti - human NR1 antibody comprising a heavy - chain variable region, a light chain, and an Fc polypeptide, wherein the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO:10, CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO:10, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO:10; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:12, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:12, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:12;

[0577] (4) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:14, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:14, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:14; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:16, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:16, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:16;

[0578] (5) A single - arm anti - human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy - chain variable region, and the heavy - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO:18, CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO:18, and CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO:18; the light chain comprises a light - chain variable region, and the light - chain variable region comprises CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO:20, CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO:20, and CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO:20.

[0579] In one embodiment, the pharmaceutical composition of the present invention comprises a single-chain anti-human NR1 antibody selected from any one of the following (1) to (5) and / or a single-chain anti-human NR1 antibody produced by post-translational modification of the single-chain anti-human NR1 antibody:

[0580] (1) A single-chain anti-human NR1 antibody comprising a heavy chain containing a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2, a light chain containing a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 4, and an Fc polypeptide;

[0581] (2) A single-chain anti-human NR1 antibody comprising a heavy chain containing a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 6, a light chain containing a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 8, and an Fc polypeptide;

[0582] (3) A single-chain anti-human NR1 antibody comprising a heavy chain containing a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10, a light chain containing a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 12, and an Fc polypeptide;

[0583] (4) A single-chain anti-human NR1 antibody comprising a heavy chain containing a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 14, a light chain containing a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 16, and an Fc polypeptide;

[0584] (5) A single-chain anti-human NR1 antibody comprising a heavy chain containing a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 18, a light chain containing a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 20, and an Fc polypeptide.

[0585] In one embodiment, the pharmaceutical composition of the present invention comprises a single-chain anti-human NR1 antibody containing a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 22, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 24, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and / or a single-chain anti-human NR1 antibody produced by post-translational modification of the single-chain anti-human NR1 antibody.

[0586] In one embodiment, the pharmaceutical composition of the present invention comprises a single-arm anti-human NR1 antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 26, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 28, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and / or a single-arm anti-human NR1 antibody produced by post-translational modification of the single-arm anti-human NR1 antibody.

[0587] In one embodiment, the pharmaceutical composition of the present invention comprises a single-arm anti-human NR1 antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and / or a single-arm anti-human NR1 antibody produced by post-translational modification of the single-arm anti-human NR1 antibody.

[0588] In one embodiment, the pharmaceutical composition of the present invention comprises a single-arm anti-human NR1 antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 34, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 36, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and / or a single-arm anti-human NR1 antibody produced by post-translational modification of the single-arm anti-human NR1 antibody.

[0589] In one embodiment, the pharmaceutical composition of the present invention comprises a single-arm anti-human NR1 antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 38, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 40, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and / or a single-arm anti-human NR1 antibody produced by post-translational modification of the single-arm anti-human NR1 antibody.

[0590] In one embodiment, the pharmaceutical composition of the present invention comprises a single-arm anti-human NR1 antibody that competitively binds to human NR1 with an anti-human NR1 antibody comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 4, and inhibits NMDAR endocytosis caused by pathogenic anti-human NR1 antibodies derived from patients with anti-NMDAR encephalitis, and / or a single-arm anti-human NR1 antibody produced by post-translational modification of the single-arm anti-human NR1 antibody.

[0591] The dosage of the single - arm anti - human NR1 antibody of the present invention varies according to the disease condition, the age of the patient, or the dosage form to be used. For example, a dosage of about 0.001 mg / kg to 100 mg / kg can be used. In addition, the dosage of the single - arm anti - human NR1 antibody of the present invention corresponding to such a dosage can be adjusted and formulated.

[0592] The pharmaceutical composition of the present invention can be used as a therapeutic agent for anti - NMDAR encephalitis.

[0593] The present invention includes a pharmaceutical composition of a single - arm anti - human NR1 antibody for treatment, which contains the single - arm anti - human NR1 antibody of the present invention and a pharmaceutically acceptable excipient. In addition, the present invention includes a method for treating anti - NMDAR encephalitis, which includes administering a therapeutically effective amount of the single - arm anti - human NR1 antibody of the present invention. In addition, the present invention contains the single - arm anti - human NR1 antibody of the present invention for treating anti - NMDAR encephalitis. In addition, the present invention includes the use of the single - arm anti - human NR1 antibody of the present invention in the manufacture of a pharmaceutical composition for treating anti - NMDAR encephalitis.

[0594] <The monovalent anti - human NR1 antibody derivative of the present invention>

[0595] In the description of the present invention, the "monovalent anti - human NR1 antibody derivative" is a monovalent antibody derivative that binds to human NR1 and contains a human NR1 antigen - binding site derived from an anti - human NR1 antibody. The monovalent anti - human NR1 antibody derivative of the present invention includes, for example, scFc, Fab fragment, F(ab')2 fragment, etc. derived from an anti - human NR1 antibody.

[0596] The monovalent anti - human NR1 antibody derivative of the present invention is a monovalent anti - human NR1 antibody derivative that contains a heavy - chain variable region and a light - chain variable region selected from any one of the following (1) to (4):

[0597] (1) A monovalent anti - human NR1 antibody derivative containing a heavy - chain variable region and a light - chain variable region, wherein the heavy - chain variable region contains CDR1 consisting of the amino - acid sequence of amino - acid numbers 31 to 35 of SEQ ID NO:6, CDR2 consisting of the amino - acid sequence of amino - acid numbers 50 to 66 of SEQ ID NO:6, and CDR3 consisting of the amino - acid sequence of amino - acid numbers 99 to 110 of SEQ ID NO:6, and the light - chain variable region contains CDR1 consisting of the amino - acid sequence of amino - acid numbers 24 to 34 of SEQ ID NO:8, CDR2 consisting of the amino - acid sequence of amino - acid numbers 50 to 56 of SEQ ID NO:8, and CDR3 consisting of the amino - acid sequence of amino - acid numbers 89 to 98 of SEQ ID NO:8;

[0598] (2) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 35 of SEQ ID NO: 10, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 66 of SEQ ID NO: 10, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 111 of SEQ ID NO: 10, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 12, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 12, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 12;

[0599] (3) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 14, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 14, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 14, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 16, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 16, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 16;

[0600] (4) A monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 31 to 36 of SEQ ID NO: 18, a CDR2 consisting of the amino acid sequence from amino acid number 51 to 66 of SEQ ID NO: 18, and a CDR3 consisting of the amino acid sequence from amino acid number 99 to 108 of SEQ ID NO: 18, and the light chain variable region comprises a CDR1 consisting of the amino acid sequence from amino acid number 24 to 34 of SEQ ID NO: 20, a CDR2 consisting of the amino acid sequence from amino acid number 50 to 56 of SEQ ID NO: 20, and a CDR3 consisting of the amino acid sequence from amino acid number 89 to 97 of SEQ ID NO: 20.

[0601] In one embodiment, the monovalent anti-human NR1 antibody derivative of the present invention is a monovalent anti-human NR1 antibody derivative comprising a heavy chain variable region and a light chain variable region selected from any one of the following (5) to (8):

[0602] (5) A monovalent anti - human NR1 antibody derivative, which comprises a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:6, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:8;

[0603] (6) A monovalent anti - human NR1 antibody derivative, which comprises a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:12;

[0604] (7) A monovalent anti - human NR1 antibody derivative, which comprises a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:14, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:16;

[0605] (8) A monovalent anti - human NR1 antibody derivative, which comprises a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:18, and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:20.

[0606] The monovalent anti - human NR1 antibody derivative of the present invention includes a monovalent anti - human NR1 antibody derivative that competitively binds to human NR1 with an anti - human NR1 antibody comprising a heavy - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:2 and a light - chain variable region consisting of the amino acid sequence shown in SEQ ID NO:4, and inhibits NMDAR internalization caused by pathogenic anti - human NR1 antibodies derived from patients with anti - NMDA receptor encephalitis. As described in the section "<The single - arm anti - human NR1 antibody of the present invention>", the monovalent anti - human NR1 antibody derivative of the present invention can be obtained by those skilled in the art.

[0607] The monovalent anti - human NR1 antibody derivative of the present invention can be modified to improve its in - vivo stability. The modifications include, for example, polyethylene glycolylation.

[0608] The present invention further includes polynucleotides encoding the antibody derivatives, expression vectors containing the polynucleotides, host cells transformed with the expression vectors, methods for producing the antibody derivatives, pharmaceutical compositions containing the antibody derivatives, the use of the antibody derivatives in the manufacture of the pharmaceutical compositions, and methods for treating patients with anti - NMDAR encephalitis with the antibody derivatives.

[0609] Specific examples are provided here to provide a further understanding of the present invention; however, these examples are provided for illustrative purposes and not for limiting the present invention.

[0610] Examples

[0611] Unless otherwise specified, the steps described in the following examples are implemented according to well-known methods. In addition, for the example parts regarding the use of commercially available kits or reagents, etc., unless otherwise specified, the experiments are carried out according to the attached protocols.

[0612] (Example 1: Obtaining antibodies from patients with anti-NMDAR encephalitis)

[0613] Based on the sequence information described in Patent Document 2 (see the description of Claim 1 and the examples in the said document), anti-human NR1 monoclonal antibodies (003-109, 007-168, 007-169, 007-124, 007-142, 008-218, P003-102, hereinafter referred to as "pathogenic anti-human NR1 antibodies") having the same heavy-chain variable region and light-chain variable region as the antibodies isolated from patients with anti-NMDAR encephalitis described in the said patent document are prepared according to conventional methods. Specifically, a polynucleotide encoding a heavy-chain polypeptide is inserted into pcDNA3.4 (Thermo Fisher Scientific) to prepare an expression vector, wherein the heavy-chain polypeptide contains a signal sequence added to the N-terminal portion of the heavy-chain variable region sequence of the pathogenic anti-human NR1 antibody, and a CH1 sequence, a hinge sequence, and an Fc sequence added to its C-terminal portion. Similarly, a polynucleotide encoding a polypeptide is inserted into pcDNA3.4 to prepare an expression vector, wherein the light-chain variable region sequence of the polynucleotide encoding the polypeptide contains a signal sequence added to the N-terminal portion of the pathogenic anti-human NR1 antibody, and a CH1 sequence added to its C-terminal portion. The expression vector is transfected into ExpiCHO-S cells (Thermo Fisher Scientific, A29127), and the antibody is obtained according to conventional methods.

[0614] The binding activities of the above seven pathogenic anti-human NR1 antibodies are evaluated by flow cytometry measurement. This flow cytometry test (Becton, Dickinson and Company, FACS Verse) is carried out in HEK cells expressing NMDAR (as described below) (the same as in Example 7). As a result, all seven pathogenic anti-human NR1 antibodies showed high binding activities in HEK cells expressing NMDAR. P003-102 (having a heavy-chain variable region composed of the amino acid sequence shown in SEQ ID NO: 2 and a light-chain variable region composed of the amino acid sequence shown in SEQ ID NO: 4) showed the highest binding activity in HEK cells expressing NMDAR (data not shown).

[0615] Note that the HEK cells expressing NMDAR used in the examples are HEK cell lines (Charles River Laboratories, CTN6121) expressing human NMDAR (NR1 / NR2B), and, unless otherwise specified, they are cultured in DMEM / F12 (Thermo Fisher Scientific, 11320033) (hereinafter referred to as "HEK cell medium expressing NMDAR") containing 10% FBS (GE Healthcare Life Sciences, SH30070.03), 50 units / mL penicillin-streptomycin (penicillin-streptomycin, Thermo Fisher Scientific, 15070063), 0.5 mg / mL geneticin (geneticin, Thermo Fisher Scientific, 10131027), 0.005 mg / mL blasticidin (blasticidin, InvivoGen, ant-bl-05), and 0.1 mg / mL zeocin (zeocin, Thermo Fisher Scientific, R25001) at 37 °C in the presence of 50% CO2.

[0616] (Example 2: Confirmation of NMDAR binding activity of the single-armed pathogenic anti-human NR1 antibody P003-102)

[0617] A monovalent anti-human NR1 antibody derivative derived from P003-102 was prepared, which showed high stability and the highest binding activity to HEK cells expressing NMDAR.

[0618] A monovalent anti - human NR1 antibody derived from P003 - 102 was prepared based on the description in the prior patent document (WO1998 / 050431). Specifically, an antibody heavy - chain expression vector was prepared, in which a polynucleotide (SEQ ID NO:21) encoding a heavy - chain polypeptide (SEQ ID NO:22) having a knobs - into - holes mutation (hereinafter referred to as "KIH mutation") in the constant region of the heavy chain of P003 - 102 was inserted into pcDNA3.4. An FC expression vector was prepared, in which an FC polypeptide (SEQ ID NO:42) - encoding polynucleotide (SEQ ID NO:41) with the KIH mutation and the above - mentioned KIH mutation was inserted into pcDNA3.4. An antibody light - chain expression vector was prepared, in which a polynucleotide (SEQ ID NO:23) encoding the light - chain polypeptide (SEQ ID NO:24) of P003 - 102 was inserted into pcDNA3.4. According to the conventional method, by transfecting three vectors, namely the antibody heavy - chain expression vector, the antibody light - chain expression vector, and the FC expression vector, into ExpiCHO - S cells and purifying the antibody molecules expressed in the cells, a monovalent antibody (single - arm antibody) (hereinafter referred to as "ABN - 5") having an antigen - binding site derived from P003 - 102 was obtained.

[0619] As described in Example 1, in HEK cells expressing NMDAR, the binding activity of ABN - 5 to human NMDAR was evaluated by flow cytometry. ABN - 5 (a single - arm antibody) showed binding activity to NMDAR expressed in HEK cells, although it was weaker than P003 - 102 (a bivalent antibody) ( Figure 1 ). In addition, ABN - 5 showed higher binding activity to human NMDAR than six other pathogenic anti - human NR1 antibodies (data not shown).

[0620] (Example 3: Internalization of NMDAR into cells by pathogenic anti - human NR1 antibodies) Pathogenic anti - human NR1 antibodies induce the internalization of NMDAR by cross - linking NMDAR on the cell surface. As a result, a decrease in the expression level of NMDAR on the cell surface has been reported (Non - Patent Document 1). The pathogenic anti - human NR1 antibody prepared in Example 1 also decreased the expression level of NMDAR on the cell surface of HEK cells.

[0621] NMDAR-expressing HEK cells were cultured overnight by replacing the NMDAR-expressing cell medium with Neurobasal medium (Thermo Fisher Scientific, 21103049) (hereinafter referred to as "induction medium") containing 10% dialyzed FBS (Thermo Fisher Scientific, 26400-044), 50 units / mL penicillin-streptomycin, 2.0 μg / mL tetracycline (tetracycline, Sigma-Aldrich, T7660), and 0.2 mM memantine (Memantine) (memantine, Sigma-Aldrich, M9292). The pathogenic anti-human NR1 antibody 007-142 at a concentration of 100 μg / mL was added. Twenty-four hours after the addition of the antibody, the expression level of NMDAR on the cell surface was evaluated by flow cytometry using a FACS Verse. Single cells were extracted by gating from the 2D extended dot plot of forward scatter light (FSC) and side scatter light (CCS) signals, followed by HEK cells or a group of cells corresponding to NMDAR-expressing HEK cells. The geometric mean fluorescence intensity (GeoMFI) of every 5000 cells in the extracted cell group was measured and used for evaluation. As a control antibody of the same isotype, an internal anti-KLH antibody (3G6) was used. When added to the medium, 007-142 reduced the expression of NMDAR on the cell surface to less than 50% (data not shown).

[0622] (Example 4: Maintaining the function of NMDAR expressed in HEK cells by pre-adding ABN-5)

[0623] The following two evaluation methods were used to determine whether ABN-5, a single-arm antibody, could rescue the reduction in the expression level of NMDAR on the cell surface and the decline in the function of NMDAR caused by the pathogenic anti-human NR1 antibody.

[0624] First, the inhibitory effect of ABN-5 on the NMDAR expression level was evaluated. Different concentrations of ABN-5 were added to the medium of NMDAR-expressing HEK cells overnight via serial dilution. Two hours after the addition, 007-142, a pathogenic human NR1 antibody, was added to the medium at a concentration of 100 μg / mL, and the expression level of NMDAR on the cell surface 24 hours after the addition of 007-142 was evaluated by flow cytometry and Example 3. 007-142 reduced the expression level of NMDAR on the cell surface of HEK cells to less than 50%. However, the NMDAR expression level on the cell surface by 007-142 was restored in a concentration-dependent manner by ABN-5 (data not shown).

[0625] To evaluate whether ABN-5 can prevent the functional decline of NMDAR through the P003-102 antibody, calcium influx was used as an index for HEK cells expressing NMDAR. Specifically, different concentrations of ABN-5 were added to the medium of HEK cells expressing NMDAR that had been cultured overnight before treatment via serial dilution. Fifteen minutes after adding ABN-5, P003-102 was added to the medium at a concentration of 0.1 μg / mL. ABN-5 was cultured for 6 hours. After removing the medium, 100 μL / well of the preparation solution of the FLIPR Calcium 6 Assay Kit (Molecular Device, R8190) was added to ABN-5 and cultured for 2 hours. NMDA (TOCRIS, 0114) with a final concentration of 30 μM was used to stimulate NMDAR. The intracellular calcium concentration was measured using the FLIPR Calcium 6 Assay Kit (Molecular Devices, R6133) by a fluorescence imaging plate reader (Molecular Devices, FLIPR TETRA). As a result, adding the P003-102 antibody to the medium led to a functional decline of NMDAR on HEK cells expressing NMDAR, and this functional decline was inhibited by ABN-5 in a concentration-dependent manner (data not shown).

[0626] As can be seen from the above, both the single-arm anti-human NR1 antibody and the pathogenic anti-human NR1 antibody bind to cell surface NMDAR. However, different from the pathogenic anti-human NR1 antibody, the single-arm anti-human NR1 antibody does not induce the internalization of NMDAR. In addition, the single-arm anti-human NR1 antibody acts antagonistically to the pathogenic anti-human NR1 antibody and prevents the functional decline of NMDAR caused by the pathogenic anti-human NR1 antibody by maintaining the expression level of NMDAR on the cell surface.

[0627] (Example 5: Isolation of a mouse antibody that binds to human NR1 and antagonistically targets the pathogenic anti-human NR1 antibody P003-102)

[0628] First, an anti-human NR1 antibody that effectively binds to P003-102 and strongly binds to human NR1 was isolated to generate a single-arm anti-human NR1 antibody with stronger binding activity to human NMDAR than ABN-5.

[0629] Specifically, mouse antibodies with high binding activity to human NR1 were isolated using BALB / cAJc1 mice and Jcl:BDF mice (both purchased from CLEA Japan). According to the conventional method, human NR1 protein was injected into the mice together with an adjuvant that triggers an immune response. The human NR1 protein for immunization was prepared by the following method.

[0630] By means of genetic engineering techniques, a linker, a HisTag sequence (His*8), an AviTag sequence (affinity), and a nucleotide sequence encoding a stop codon were added to the 3'-end of the polynucleotide encoding the extracellular domain of human NR1 (SEQ ID NO: 43, the corresponding amino acid sequence is shown in SEQ ID NO: 44), and inserted into pcDNA3.1 (Thermo Fisher Scientific), and became an expression vector for a fusion protein of human NR1 and HisTag (hereinafter referred to as "human NR1-His protein"). The expression vector was transfected into Expi293F cells (Thermo Fisher Scientific, A14528). The human NR1-His protein was purified from the culture supernatant.

[0631] After injecting mice several times with the purified human NR1-His protein and an adjuvant according to a conventional method, lymphocytes were collected from the spleen or lymph nodes of the mice.

[0632] The human NR1-His protein was fluorescently labeled using an Alexa Fluor 647 antibody labeling kit (Thermo Fisher Scientific, A20186). Based on the description in a prior non-patent literature (BMC Biol., 2012, Vol. 10, Article No. 80), B cells that bound to the fluorescently labeled human NR1-His protein were sorted from mouse lymphocytes using a cell sorter (Becton Dickinson, FACSmelody). According to the method, the nucleic acid encoding the antibody extracted from the sorted B cells was cloned and used to produce a mouse antibody that binds to human NR1. The human NR1-His protein was used, and an ELISA binding assay was performed according to a conventional method (the specific conditions are the same as those for the following ELISA competitive assay) to evaluate the NR1 binding activity of the produced mouse antibody. As a result, a large number of mouse antibodies with higher binding activity to human NR1 than the P003-102 antibody could be isolated (data not shown).

[0633] An ELISA competitive assay was performed to determine which antibody has antagonistic activity against P003-102 from isolated mice. Specifically, 15 μL of human NR1-His protein solution (1 μg / mL, in Tris-buffered saline (referred to herein as "TBS")) was added to a 384-well plate (Thermo Fisher Scientific, 460372) and incubated overnight at 4°C. Then, the human NR1-His protein solution was removed and washed with 0.5% Tween-20 (NIPPON GENE, 310-07375) containing TBS (referred to as "TBS-T"). After washing, 50 μL of Blocking One (Nacalai Tesque, Inc., 03953-95) was added and incubated at room temperature for 1 hour or longer. The blocking solution was removed and washed as described above. 15 μL of mouse anti-human NR1 antibody at a concentration of 100,000 ng / mL to 0.1 ng / mL (TBS-T containing 5% BlockingOne) or a control antibody (P003-102) was added to each well and incubated at room temperature for 10 to 20 minutes.

[0634] P003-102 was biotinylated using a Biotin Labeling Kit-NH2 (DOJINDO and LK03). 15 μL of biotinylated P003-102 (600 ng / mL, in a dilution solution) was added and incubated at room temperature for 1 hour. After washing, 15 μL was added to HRP-labeled streptavidin (Funakoshi Co., Ltd., 21130) diluted 8,000-fold with a diluent and incubated at room temperature for 30 minutes. After washing, 15 μL was added to BM Chemiluminescence ELISA Substrate (POD) (Roche Diagnotics 11 582 950001, which is a chemiluminescence detection reagent for measuring chemiluminescence in a multi-label plate reader (PerkinElmer, 2103 EnVision)). The inhibition rate ranged from 100% for the 100,000 ng / mL measurement of the control antibody to 0% for the 0 ng / mL measurement. The inhibition rate was analyzed by four-parameter logistic curve fitting. The inhibition rate of each sample was calculated using the following formula.

[0635] Inhibition rate (%) = (1 - (Sample measurement value - 100% measurement value) / (0% measurement value - 100% measurement value)) x 100, and samples with a negative inhibition rate were considered 0%.

[0636] Analyzed were anti-mouse anti-human NR1 monoclonal antibodies having antagonistic activity against biotin-labeled P003-102. Among the anti-mouse anti-human NR1 antibodies, ABN_1-175, ABN_1-178.3, ABN_3-385, and ABN_3-502 showed strong antagonistic activity against P003-102. In particular, ABN_1-178.3 and ABN_3-502 had the strongest antagonistic activity against P003-102( Figure 2 ).

[0637] (Example 6: Method for Producing the Single-Arm Anti-Human NR1 Antibody of the Present Invention) Since four mouse anti-NR1 monoclonal antibodies (ABN_1-175, ABN_1-178.3, ABN_3-385, and ABN_3-502) showed strong antagonistic activity against P003-102, humanized single-arm anti-human NR1 antibodies were produced.

[0638] The amino acid sequences of the heavy chain variable region (VH) and the light chain variable region (VL) of ABN_1-175, ABN_1-178.3, ABN_3-385, or ABN_3-502 were designed using the techniques described in a prior non-patent literature (Biosci., 2008, Vol. 13, pp. 1619-1633). Based on well-known techniques, human antibody framework sequences were multiply selected from the germline framework database, and the human antibody framework sequences had amino acid sequences highly homologous to the amino acid sequences of the VH and VL framework regions (FR) of the mouse antibody based on the template human antibody, as the transplantation sites for the complementarity determining region (CDR) amino acids in VH and VL. Using this sequence information, the amino acid sequences of multiple humanized antibodies from one mouse antibody were designed. The CDR sequences were determined from the amino acid sequences obtained in Example 5, with reference to the Kabat database ("Sequences of Proteins of Immunological Interest", US Department of Health and Human Services, US Government Printing Office). In addition, ABN-5 (Example 2) was a multiple of humanized single-arm anti-human NR1 monoclonal antibodies from each mouse produced by combining with the constant region sequence having the KIH mutant, respectively. These humanized single-arm anti-human NR1 monoclonal antibodies were single-arm antibodies composed of one heavy chain polypeptide, one light chain polypeptide, and one Fc polypeptide of any one of humanized ABN_1-175, ABN_1-178.3, ABN_3-385, and ABN_3-502.

[0639] (Example 7: Measuring the NMDAR Binding Activity of a Humanized Single-Arm Anti-Human NR1 Antibody)

[0640] Using HEK cells expressing NMDAR, the binding activity of the generated humanized single-arm anti-human NR1 monoclonal antibody was evaluated in a flow cytometry assay.

[0641] The HEK cells expressing NMDAR were induced overnight and collected. After recovery, per 1.5x10 5 cells were counted, and the humanized single-arm anti-human NR1 antibody and ABN-5 derived from a mouse antibody were added at an initial concentration of 30 μg / mL and serially diluted 3-fold to generate 11 different concentrations, and incubated with the cells at 4 °C for 30 minutes. After staining, the cells were pelleted at 1,200 rpm for 5 minutes and the supernatant was removed. After adding 2% dialyzed FBS to the cells, the cells were centrifuged and pelleted. After removing the supernatant, the PE-labeled anti-human IgG antibody (Jackson ImmunoResearch Laboratories, 109-116-098) was diluted 100-fold with 2% dialyzed FBS and added to the cells at 4 °C for 30 minutes. After staining, the cells were centrifuged and pelleted as described above. After removing the supernatant, the cells were resuspended in 2% dialyzed FBS, and the steps after secondary staining were repeated. Flow cytometry (FACS Verse) was measured as described in Example 3. Using FlowJo (Becton Dickinson, version 10.0.8), each antibody that binds to cell surface NMDAR was analyzed by GeoMFI showing the PE fluorescence intensity value.

[0642] The results were consistent that multiple humanized single-arm anti-human NR1 antibodies generated in Example 6 bound more strongly to NMDAR than the P003-102 antibody. For example, compared with ABN-5, three types of humanized single-arm anti-human NR1 antibodies (ABN_178-m1, ABN_178-m2, ABN_178-m3) derived from the mouse antibody ABN_1 -178.3 bound very strongly to NMDAR ( Figure 3 ).

[0643] Representative humanized single-chain anti-human NR1 antibodies are selected from the murine anti-human NR1 antibodies ABN_1-1-175, ABN_1-1-178.3, ABN_3-385, ABN_1-178.3 and ABN_3-385, and ABN_3-502, and are named ABN-1, ABN-2 (the same antibody as ABN_178-m3 above), ABN-3 and ABN-4, respectively. The amino acid sequences and nucleotide sequences of the VH, VL, heavy chain, light chain and Fc polypeptides of ABN-5 described in Example 2 and each newly generated humanized single-chain anti-human NR1 antibody are shown in Table 1 (in Table 1, the heavy chain, light chain, Fc polypeptide, amino acid sequence and nucleotide sequence are abbreviated as HC, LC, Fc, AA and NA, respectively).

[0644] Table 1

[0645]

[0646]

[0647] The CDR1, CDR2 and CDR3 of VH shown in SEQ ID NO:2 are respectively composed of the amino acid sequences of amino acid numbers 26 to 34, 52 to 58 and 97 to 109 of SEQ ID NO:2. The CDR1, CDR2 and CDR3 of VL shown in SEQ ID NO:4 are respectively composed of the amino acid sequences of amino acid numbers 26 to 33, 51 to 53 and 92 to 100 of SEQ ID NO:4.

[0648] The CDR1, CDR2 and CDR3 of VH shown in SEQ ID NO:6 are respectively composed of the amino acid sequences of amino acid numbers 31 to 35, 50 to 66 and 99 to 110 of SEQ ID NO:6. The CDR1, CDR2 and CDR3 of VL shown in SEQ ID NO:8 are respectively composed of the amino acid sequences of amino acid numbers 24 to 34, 50 to 56 and 89 to 98 of SEQ ID NO:8.

[0649] The CDR1, CDR2 and CDR3 of VH shown in SEQ ID NO:10 are respectively composed of the amino acid sequences of amino acid numbers 31 to 35, 50 to 66 and 99 to 111 of SEQ ID NO:10. The CDR1, CDR2 and CDR3 of VL shown in SEQ ID NO:12 are respectively composed of the amino acid sequences of amino acid numbers 24 to 34, 50 to 56 and 89 to 97 of SEQ ID NO:12.

[0650] The CDR1, CDR2, and CDR3 of VH shown in SEQ ID NO:14 are composed of the amino acid sequences from amino acid numbers 31 to 36, 51 to 66, and 99 to 108 of SEQ ID NO:14, respectively. The CDR1, CDR2, and CDR3 of VL shown in SEQ ID NO:16 are composed of the amino acid sequences from amino acid numbers 24 to 34, 50 to 56, and 89 to 97 of SEQ ID NO:16, respectively.

[0651] The CDR1, CDR2, and CDR3 of VH shown in SEQ ID NO:18 are composed of the amino acid sequences from amino acid numbers 31 to 36, 51 to 66, and 99 to 108 of SEQ ID NO:18, respectively. The CDR1, CDR2, and CDR3 of VL shown in SEQ ID NO:20 are composed of the amino acid sequences from amino acid numbers 24 to 34, 50 to 56, and 89 to 97 of SEQ ID NO:20, respectively.

[0652] To evaluate the binding activities of ABN-2, ABN-4, and ABN-5 to human NMDAR, surface plasmon resonance (SPR) analysis was performed. Biacore T200 (GE Healthcare) was used for this SPR analysis.

[0653] The human antibody capture kit (GE Healthcare, BR-1008-39) was immobilized onto a CM5 sensor chip (GE Healthcare, BR-1005-30) and an amine coupling kit (GE Healthcare, BR-1000-50) using anti-human IgG (human antibody capture kit).

[0654] The 1st flow channel was used as a reference, and no humanized single-arm anti-human NR1 antibody bound to this flow channel. Humanized anti-human NR1 antibodies with concentrations of 2, 5, and 10 μg / mL were prepared in HBS-EP+ buffer (GE Healthcare, BR-1006-69), added at a flow rate of 10 μL / min for 50 seconds in another stream (the 2nd, 3rd, and 4th streams), and the antibodies were immobilized. Then, human NR1-His protein diluted to 0.5 μg / mL in HBS-EP+ buffer was added at a flow rate of 50 μL / min within 2 minutes. The binding of the humanized single-arm anti-human NR1 antibody and humanized NR1-His protein was measured. After adding HBS-EP+ buffer at a flow rate of 50 μL / min for 5 minutes, the dissociation of the humanized single-arm anti-human NR1 antibody and human NR1-His protein was measured. The association rate constant (ka) and dissociation rate constant (kd) were analyzed and calculated using a 1:1 binding model, and the dissociation constant (KD) was calculated by dividing kd by ka. Finally, for the KD value of the humanized single-arm anti-human NR1 antibody, the KD value of 10 μL / mL was used under the solid-phase conditions of this antibody.

[0655] As a result of the SPR analysis, the KD (M) of ABN-2 was 6.92E-10, which had the highest binding activity against human NR1-His protein among ABN-2, ABN-4, and ABN-5. The KD values of ABN-2, ABN-4, and ABN-5 against human NR1-His protein are shown in Figure 2 . The KD values of ABN-2 and ABN-4 were much smaller than the KD value of ABN-5.

[0656] Table 2

[0657] ABN-2 ABN-4 ABN-5 KD(M) 6.92E-10 8.78E-10 1.97E-07

[0658] (Example 8: Competitive binding activities of ABN-2, ABN-4, and ABN-5 to P003-102)

[0659] In a similar manner to Example 5, the competitive binding activities of ABN-2, ABN-4, and ABN-5 against human NR1 to P003-102 were measured by ELISA competitive assay. The results showed that in terms of binding to NMDAR, the antagonistic activity of ABN-2 against P003-102 was stronger than that of ABN-4 and ABN-5. ( Figure 4 ).

[0660] (Example 9: Restoration of the cell surface expression level of NMDAR by adding ABN-2, ABN-4, and ABN-5)

[0661] To evaluate the inhibitory effect of ABN-2 and ABN-5 on the internalization of cell surface NMDAR caused by pathogenic anti-human NR1 antibodies such as P003-102 (inhibiting NMDAR internalization), flow cytometry was used to evaluate NMDAR internalization.

[0662] Using the induction medium, a pathogenic anti-human NR1 antibody mixture for stimulating NMDAR internalization was prepared at the concentrations shown in Table 3.

[0663] Table 3

[0664]

[0665] The pathogenic anti-human NR1 antibody mixture was added to a 96-well plate (IWAKI, 3860-096) at 20 μL / well. HEK cells expressing NMDAR were prepared at 2.5x10 6 cells / mL in the induction medium and inoculated at 60 μL / well, and cultured overnight under the conditions of 37 °C and 5% CO2. Serial dilutions of ABN-2 or ABN-5 at 20 μL / well were added and incubated overnight, diluted 3-fold serially to generate 10 different measurement points. Then, the recovered cells were treated with ABN-2 in the ABN-2 treatment group and P003-102 in the ABN-5 treatment group, and primary staining was performed at 5.0 μg / mL each for 30 minutes at 4 °C. After staining, the cells were pelleted at 1,200 rpm for 5 minutes. The supernatant was removed, and 2% dialyzed FBS containing D-PBS(-) was added to the cells. After adding D-PBS, the cells were centrifuged and the supernatant was removed. A PE-labeled anti-human IgG antibody diluted 100-fold with 2% dialyzed FBS containing D-PBS(-) was used for secondary staining at 4 °C for 30 minutes. After staining, the cells were centrifuged and the supernatant was removed as described above. The cells were suspended in 2% dialyzed FBS containing D-PBS(-), and the steps after secondary staining were repeated. As described in Example 3, flow cytometry was measured using FACA Verse. Using flow cytometry analysis software FlowJo, the PE fluorescence intensity value was determined by GeoMFI to analyze the NMDAR expression level on the cell surface.

[0666] The NMDAR expression level on the cell surface was evaluated as the cell surface NR1 expression rate. The cell surface NR1 expression rate used the PE fluorescence intensity of HEK cells expressing NMDAR as an index, and the relative value was calculated based on the 100% value when stained with ABN-2 and P003-102 under the condition of no pathogenic anti-human NR1 antibody mixture and the 0% value when stained with an isotype control antibody (3G6, an internally generated anti-KLH antibody).

[0667] When using ABN-2 and ABN-5, the cell surface NR1 expression rate is shown in Figure 5 For the pathogenic anti-human NR1 antibody-dependent reduction of the cell surface NMDAR expression level, ABN-2 showed a much higher recovery effect than ABN-5. For the pathogenic anti-human NR1 antibody-dependent reduction of the cell surface NMDAR expression level, ABN-4 also showed a high recovery effect, although lower than ABN-2 (data not shown).

[0668] (Example 10: Evaluation of the restoration of NMDAR function by ABN-2 and ABN-4) The cell calcium concentration was measured to evaluate the effects of ABN-2 and ABN-4 against the NMDAR hypofunction caused by the pathogenic antibody P003-102.

[0669] 100 uL of HEK cells expressing NMDAR were seeded at 1.5 x 105 cells / well in a poly-D-lysine-coated 96-well plate (Corning, 356640) in the assay medium (NeuroBasal medium containing 0.2 mM memantine and 10% FBS). After incubation overnight at 37 °C and 5% CO2, 25 uL of P003-102 diluted to a final concentration of 1 μg / mL in the above assay medium was added to each well. After incubation for 15 minutes at 37 °C and 5% CO2, 25 uL of ABN-2 diluted to a final concentration of 10 to 0.01 μg / mL was added to each well. After incubation for 6 hours at 37 °C and 5% CO2, the medium was removed. As described in Example 4, 100 ul of the prepared solution of the FLIPR Calcium 6 Assay Kit (Molecular Device, R8190) was added to each well and incubated for 2 hours at 37 °C and 5% CO2. Ca 2+ influx was measured using FLIPR TETRA. Ca 2+ influx was used as an index to evaluate the function of NMDAR.

[0670] The results showed that ABN-2 had a dose-dependent inhibitory effect on P003-102-induced NMDAR hypofunction ( Figure 6 ). The same test was also performed using ABN-4 and ABN-5. ABN-4 showed a weaker inhibitory effect than ABN-2 (data not shown).

[0671] (Example 11: Preparation of P003-102 and ABN-2 for animal testing)

[0672] Prepare P003-102 and ABN-2 used in Example 12.

[0673] Prepare the expression vector for P003-102. For the heavy chain expression of P003-102, a polynucleotide (SEQ ID NO:45) encoding a heavy chain polypeptide (SEQ ID NO:46) consisting of the heavy chain variable region sequence (SEQ ID NO:2), CH1 sequence, hinge sequence and FC sequence of P003-102 was ligated to the 3'-side of a polynucleotide encoding a signal sequence (MEWSWVFLFFLSVTTGVHS; SEQ ID NO:49), and the ligated polynucleotide was inserted into the GS vector pEE6.4 (Lonza). In addition, for the light chain expression of P003-102, a polynucleotide (SEQ ID NO:47) encoding a polypeptide (SEQ ID NO:48) in which the light chain variable region sequence (SEQ ID NO:4) and CL sequence were added to the 3'-side of a polynucleotide encoding a signal sequence (MSVPTQVLGLLLLWLTDARC; SEQ ID NO:50) was ligated, and the ligated polynucleotide was inserted into the GS vector pEE12.4 (Lonza). These GS vectors were treated with restriction enzymes NotI-HF (New England Biolabs, R3189L) and PvuI-HF (New England Biolabs, R3150L). Ligation of these restriction enzyme-treated fragments was carried out using a DNA ligation kit (Ligation-Convenience Kit, NIPPONGENE, 319-05961). A dual GS vector (DGV) containing two genes of the heavy chain and light chain was constructed. Transfection of the DGV into CHO-K1SV cells (Lonza) was performed, and a stable cell line expressing P003-102 was prepared. P003-102 was purified from the culture supernatant of the cells according to a conventional method.

[0674] For the heavy chain expression of ABN-2, a polynucleotide (SEQ ID NO:29) encoding a heavy chain polypeptide (SEQ ID NO:30) was ligated to which a heavy chain variable region sequence (SEQ ID NO:10) of ABN-2, a CH1 sequence, a hinge sequence, and an FC sequence with KIH mutation were added to the 3'-side of a polynucleotide encoding a signal sequence (MEWSWVFLFFLSVTTGVHS; SEQ ID NO:49), and the ligated polynucleotide was inserted into the GS vector pEE6.4. In addition, for the light chain expression of ABN-2, a polynucleotide (SEQ ID NO:31) encoding a polypeptide (SEQ ID NO:32) to which a light chain variable region sequence (SEQ ID NO:12) and a CH1 sequence were added to the 3'-side of a polynucleotide encoding a signal sequence (MSVPTQVLGLLLLWLTDARC; SEQ ID NO:50) was inserted into the GS vector pEE12.4. These GS vectors were subjected to restriction enzyme treatment with NotI-HF and PvuI-HF. The ligations of these restriction enzyme-treated fragments were performed with Mighty Mix, and a dual GS vector (DGV) containing two genes of the heavy chain and the light chain was constructed. In addition, for the expression of an FC polypeptide (SEQ ID NO:42) with a KIH mutation paired with the KIH mutation of the heavy chain polypeptide, a polynucleotide (SEQ ID NO:41) encoding the FC polypeptide was ligated to the 3'-side of a polynucleotide encoding a signal sequence (MEWSWVFLFFLSVTTGVHS; SEQ ID NO:49), and the ligated polynucleotide was inserted into the GS vector pEE 12.4. Transfection of two types of pEE12.4 expression vectors containing polynucleotides encoding the DGV and the Fc polypeptide was performed, and a stable cell line expressing ABN-2 was prepared. ABN-2 was purified from the culture supernatant of the cells according to a conventional method.

[0675] (Example 12: Preparation of a marmoset model of anti-NMDAR encephalitis-like and evaluation of the therapeutic effect of ABN-2 on the marmoset model)

[0676] An anti-NMDAR encephalitis model was prepared using common marmosets (hereinafter referred to as "marmosets"), and the therapeutic effect of ABN-2 was evaluated in the model. As described in Example 11, the model was prepared by passive immunization with P003-102. For the infusion of P003-102, under isoflurane anesthesia (Mylan Inc., 871119), the marmoset was fixed to a stereotaxic apparatus (NARISHIGE, SR-6C-HT) with a ventilator (SHINANO, SN-480-7). A cannula (Plastics One (registered trademark), 813220PSPCLC) was placed at the target in the cerebral ventricle (A / P: 3.5 mm, M / L: ±0.0 mm, Dept: 6.95 mm (from the dura mater)). Regarding the target for placing the cannula, a marmoset brain atlas (stereotaxic map of the marmoset brain, NCBI Bookshelf ID: NBK55612) was used as a reference. Next, the cannula in the cerebral ventricle was connected to a microinfusion pump (PRIMETECH, iPRECIO (registered trademark) SMP-200). The pump was set at a jacket pocket (inside) where behavior observation was not affected. After placing the cannula in the cerebral ventricle, PBS (FUJIFILM Wako Pure Chemical Corporation, 045-29795) was allowed to flow out from the microinfusion pump at a flow rate of 2 μL / h for one week. Subsequently, the pump was replaced with a microinfusion pump containing P003-102 (5,000 μg / mL), and P003-102 was infused into the cerebral ventricle at a flow rate of 2 μL / h (10 μg / h). This time point was set as Day 0 of administration. The single treatment with P003-102 was carried out for 14 days. Subsequently, the pump was replaced with a microinfusion pump containing a mixture of an equal amount of P003-102 and a therapeutic antibody (ABN-2) (each 2,500 μg / mL). The two types of antibodies were infused into the cerebral ventricle at a flow rate of 4 μL / h (each 10 μg / h) for 14 days. The group of treated marmosets was called the "ABN-2 administration group" (n = 9). The group of marmosets treated with a control antibody (3G6, which is an internal anti-KLH antibody) was called the "control antibody administration group" (n = 3). The following observations were made.

[0677] Method for observing abnormal behavior

[0678] Before administering P003-102 into the cerebral ventricle (day 0), on the 14th day after starting the administration of P003-102 (day 14), and on the 14th day after administering ABN-2 or control antibody together with P003-102 (day 28), videos of approximately 5 minutes were recorded with a camera at a predetermined time. Based on the videos, the abnormal behavior score was evaluated using the marmoset version of the Abnormal Rating Scale (Abnormal Rating Scale: ARS), which is a modified Unified Parkinson's Disease Rating Scale (UPDRS, Behav Brain Res., 2008, Vol. 194, pp. 152-161). This evaluation was performed by a person experienced in observing the behavior of marmosets under blind conditions. ARS is a scoring method with a maximum score of 22 points, which uses a list of psychological abnormalities (attention, motivation, fear / anxiety) and a list of movement disorders (speed of voluntary movement, coordination of movement, presence or absence of jumping, stereotyped behavior, abnormalities of the arms and legs) as evaluation indicators.

[0679] Evaluating abnormal behavior with ARS:

[0680] It is recognized that since P003-102 was administered into the cerebral ventricle of marmosets, the ARS value increased (data not shown). Compared with day 0, the ARS value (expression of abnormal behavior) increased significantly on day 14 (in the middle of Figure 7 ). In the ABN-2 administration group, a significant decrease in the ARS value was observed on day 28. On the other hand, the control antibody administration group also showed a high ARS value even on day 28 (on the right side of Figure 7 ). Compared with the control antibody administration group, the ABN-2 administration group showed a significant improvement in the abnormal behavior induced by P003-102 (indicated by the decrease in the ARS value). Figure 8 The transition of ARS of each individual marmoset in the ABN-2 administration group on day 0, day 14, and day 28 is shown.

[0681] It is expected that the monovalent anti-human NR1 antibody derivative of the present invention can be used for the treatment of anti-NMDAR encephalitis. In addition, the methods for producing the polynucleotide, expression vector, transformed host cell, and antibody of the present invention can be used to produce anti-human NR1 antibody derivatives.

[0682] Sequence Listing Plain Text

[0683] In the case of “artificial sequences”, the description is given in the numerical heading <223> in the following Sequence Listing. Specifically, the sequences shown in SEQ ID NO:1 and 3 in the Sequence Listing are codon-optimized nucleotide sequences encoding the heavy chain variable region and the light chain variable region of ABN-5. The nucleotide sequences shown in SEQ ID NO:5 and 7 encode the heavy chain variable region and the light chain variable region of ABN-1. The nucleotide sequences shown in SEQ ID NO:9 and 11 in the Sequence Listing encode the heavy chain variable region and the light chain variable region of ABN-2. The nucleotide sequences shown in SEQ ID NO:13 and 15 in the Sequence Listing encode the heavy chain variable region and the light chain variable region of ABN-3. The nucleotide sequences shown in SEQ ID NO:17 and 19 in the Sequence Listing encode the heavy chain variable region and the light chain variable region of ABN-4. Each amino acid sequence shown in SEQ ID NO:6, 8, 10, 12, 14, 16, 18 or 20 is the amino acid sequence of the heavy chain variable region or the light chain variable region encoded by the nucleotide sequence shown in SEQ ID NO:5, 7, 9, 11, 13, 15, 17 or 19, respectively. The nucleotide sequences shown in SEQ ID NO:21 and 23 encode the heavy chain and the light chain of ABN-5. The nucleotide sequences shown in SEQ ID NO:25 and 27 encode the heavy chain and the light chain of ABN-1. The nucleotide sequences shown in SEQ ID NO:29 and 31 are nucleotide sequences encoding the heavy chain and the light chain of ABN-2. The nucleotide sequences shown in SEQ ID NO:33 and 35 encode the heavy chain and the light chain of ABN-3. The nucleotide sequences shown in SEQ ID NO:37 and 39 encode the heavy chain and the light chain of ABN-4. Each amino acid sequence shown in SEQ ID NO:22, 24, 26, 28, 30, 32, 34, 36, 38 or 40 is the amino acid sequence of the heavy chain or the light chain encoded by the nucleotide sequence shown in SEQ ID NO:21, 23, 25, 27, 29, 31, 33, 35, 37 or 39, respectively. The nucleotide sequence shown in SEQ ID NO:41 encodes the Fc polypeptide of ABN-1-5, and the amino acid sequence shown in SEQ ID NO:42 is the amino acid sequence encoded by the nucleotide sequence shown in SEQ ID NO:41. The nucleotide sequence shown in SEQ ID NO:43 encodes the extracellular domain of human NR1 for producing human NR1-His protein in Example 5. The amino acid sequence shown in SEQ ID NO:46 is the amino acid sequence of the heavy chain of P003-102 encoded by the nucleotide sequence shown in SEQ ID NO:45. The amino acid sequence shown in SEQ ID NO:48 is the amino acid sequence of the light chain of P003-102 encoded by the nucleotide sequence shown in SEQ ID NO:47.The amino acid sequence shown in SEQ ID NO:49 is the signal sequence for expressing the heavy chains of P003-12 and ABN-2 and the Fc polypeptide used in Example 11. The amino acid sequence shown in SEQ ID NO:50 is the signal sequence for expressing the light chains of P003-12 and ABN-2 used in Example 11. Sequence Listing <110> Ariad Pharmaceuticals, Inc. <120> Anti-human NR1 antibody derivative <130> A21009A00 <150> JP2020-092105 <151> 2020-05-27 <160> 50 <170> PatentIn version 3.5 <210> 1 <211> 360 <212> DNA <213> Artificial Sequence <220> <223> Codon-optimized <220> <221> CDS <222> (1)..(360) <400> 1 cag gtc cag ctg caa gag tct ggc cct gga ctg gtc aag cct tct ggc 48 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly 1 5 10 15 acc ctg tct ctg aca tgt gct gtg tcc ggc ggc tcc atc tcc tcc tct 96 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser 20 25 30 aat tgg tgg tct tgg gtc cga cag cct cct ggc aaa gga ctg gaa tgg 144 Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 atc ggc gag atc tac cac tcc ggc aac acc aac tac aac ccc agc ctg 192 Ile Gly Glu Ile Tyr His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu 50 55 60 aag tcc aga gtg acc gtg tcc gtg gac aag tcc aag aac cag ttc tcc 240 Lys Ser Arg Val Thr Val Ser Val Asp Lys Ser Lys Asn Gln Phe Ser 65 70 75 80 ctg aag ctg acc tct gtg acc gct gcc gat acc gcc gtg tac tac tgt 288 Leu Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 gct aga gat gtg tct ggc gga gtg aat tgg ttc gat cct tgg ggc cag 336 Ala Arg Asp Val Ser Gly Gly Val Asn Trp Phe Asp Pro Trp Gly Gln 100 105 110 ggc aca ctg gtt acc gtg tcc tct 360 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 2 <211> 120 <212> PRT <213> Homo sapiens <400> 2 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser 20 25 30 Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Glu Ile Tyr His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Val Ser Val Asp Lys Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Thr Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Val Ser Gly Gly Val Asn Trp Phe Asp Pro Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 3 <211> 330 <212> DNA <213> Artificial Sequence <220> <223> Codon Optimized <220> <221> CDS <222> (1)..(330) <400> 3 aac ttt atg ctg acc cag cct cac tcc gtg tcc gag tct cca ggc aag 48 Asn Phe Met Leu Thr Gln Pro His Ser Val Ser Glu Ser Pro Gly Lys 1 5 10 15 acc gtg acc atc tcc tgt acc aga tcc tcc ggc tct atc gcc tcc aac 96 Thr Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ser Ile Ala Ser Asn 20 25 30 tac gtg cag tgg tat cag cag agg cct ggc tct gct cct acc acc gtg 144 Tyr Val Gln Trp Tyr Gln Gln Arg Pro Gly Ser Ala Pro Thr Thr Val 35 40 45 atc tac gag gac aac cag agg cct tct ggc gtg ccc gat agg ttc tct 192 Ile Tyr Glu Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 ggc tcc atc gac tcc tct tcc aac tcc gcc tct ctg acc atc agc ggc 240 Gly Ser Ile Asp Ser Ser Ser Asn Ser Ala Ser Leu Thr Ile Ser Gly 65 70 75 80 ctg aaa acc gag gac gag gcc gac tac tac tgc cag tcc tac gac tct 288 Leu Lys Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser 85 90 95 tcc acc gtg gtg ttt ggc ggc gga aca aag ctg aca gtg ctg 330 Ser Thr Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 <210> 4 <211> 110 <212> PRT <213> Homo sapiens <400> 4 Asn Phe Met Leu Thr Gln Pro His Ser Val Ser Glu Ser Pro Gly Lys 1 5 10 15 Thr Val Thr Ile Ser Cys Thr Arg Ser Ser Gly Ser Ile Ala Ser Asn 20 25 30 Tyr Val Gln Trp Tyr Gln Gln Arg Pro Gly Ser Ala Pro Thr Thr Val 35 40 45 Ile Tyr Glu Asp Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Ile Asp Ser Ser Ser Asn Ser Ala Ser Leu Thr Ile Ser Gly 65 70 75 80 Leu Lys Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser 85 90 95 Ser Thr Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 <210> 5 <211> 363 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(363) <400> 5 gaa gtt cag ctg gtt cag tct ggc gcc gaa gtg aag aag cct ggc gag 48 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 tcc ctg aag atc tcc tgc aaa ggc tcc ggc tac acc tac acc agc tac 96 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Tyr Thr Ser Tyr 20 25 30 tgg atg aac tgg gtc cga cag atg cct ggc aaa ggc ctg gaa tgg atg 144 Trp Met Asn Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 ggc aga atc gac ccc tac gac tcc gag aca cac tac gac cag aaa ttc 192 Gly Arg Ile Asp Pro Tyr Asp Ser Glu Thr His Tyr Asp Gln Lys Phe 50 55 60 aag gac caa gtg acc ctg agc gtg gac aag tcc atc tcc acc gct tac 240 Lys Asp Gln Val Thr Leu Ser Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 ctg cag tgg tcc tct ctg aag gcc tct gac acc gcc atg tac tac tgt 288 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 gct ggc ggc atc acc acc atc ctc ggc ggc tac ttt gat tac tgg ggc 336 Ala Gly Gly Ile Thr Thr Ile Leu Gly Gly Tyr Phe Asp Tyr Trp Gly 100 105 110 cag ggc aca ctg gtc acc gtt tct tcc 363 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 6 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 6 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Tyr Thr Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Tyr Asp Ser Glu Thr His Tyr Asp Gln Lys Phe 50 55 60 Lys Asp Gln Val Thr Leu Ser Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Gly Gly Ile Thr Thr Ile Leu Gly Gly Tyr Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 7 <211> 327 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(327) <400> 7 gac atc cag atg acc cag tct cca tcc tct ctg tcc gcc tct gtg ggc 48 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 gac aga gtg acc atc acc tgt aga gcc agc cag gac atc tcc aac tac 96 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr 20 25 30 ctg aac tgg tat cag cag aag ccc ggc aag gcc cct aag ctg ctg atc 144 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 tac tac acc tct cgg ctg cac tct ggc gtg ccc tct aga ttt tct ggc 192 Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 tcc ggc tct ggc acc gac tat acc ctg aca atc tcc agc ctg cag cct 240 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 gag gac ttc gcc acc tac tat tgc cag cag ggc aac acc ctg cct cca 288 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Pro 85 90 95 tac aca ttt ggc cag ggc acc aag gtg gaa atc aag cgt 327 Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 <210> 8 <211> 109 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 8 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Thr Leu Pro Pro 85 90 95 Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 <210> 9 <211> 366 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(366) <400> 9 gaa gtg cag ctg gtt gaa tct ggc ggc gga ttg gtt cag cct ggc gga 48 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 tct ctg aga ctg tcc tgc aag gct tct ggc tac gcc tac acc tcc tac 96 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Tyr Ala Tyr Thr Ser Tyr 20 25 30 tgg atg aac tgg gtc cga cag gct cct ggc aaa gga ctc gtg tgg gtc 144 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45 gga aga atc gac ccc tac gac tcc gag aca cac tac aac cag aag ttc 192 Gly Arg Ile Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gln Lys Phe 50 55 60 aag gac cgg ttc acc ctg agc gtg gac aag gcc aag tct acc acc tac 240 Lys Asp Arg Phe Thr Leu Ser Val Asp Lys Ala Lys Ser Thr Thr Tyr 65 70 75 80 ctg cag atg aac tcc ctg aga gcc gag gac acc gcc gtg tac tat tgc 288 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 gct acc ctg atc acc aca ctg aga ggc gag ggc gcc atg gaa tat tgg 336 Ala Thr Leu Ile Thr Thr Leu Arg Gly Glu Gly Ala Met Glu Tyr Trp 100 105 110 gga cag gga acc ctg gtc acc gtg tcc tct 366 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 10 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 10 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Ala Ser Gly Tyr Ala Tyr Thr Ser Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Val Trp Val 35 40 45 Gly Arg Ile Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Arg Phe Thr Leu Ser Val Asp Lys Ala Lys Ser Thr Thr Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Leu Ile Thr Thr Leu Arg Gly Glu Gly Ala Met Glu Tyr Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 11 <211> 324 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(324) <400> 11 gac ctg cag atg acc cag tct cct tcc agc ctg tct gcc tct gtg ggc 48 Asp Leu Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 gac aga gtg acc atc aca tgc aag gcc agc cag gac gtg aac acc gcc 96 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 gtt gct tgg tat cag cag aag cct ggc aag gcc cct aag ctg ctg atc 144 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 tac tgg gcc tcc acc aga cat acc ggc gtg ccc tct aga ttc tcc ggc 192 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 tct ggc tct ggc acc gac tat acc ctg aca atc tcc agc ctg cag cct 240 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 gag gac ttc gcc acc tac tac tgc cag cag cac tac agc acc cct ttc 288 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Phe 85 90 95 acc ttt ggc cag ggc acc aag gtg gaa atc aag cgt 324 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 <210> 12 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 12 Asp Leu Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Ser Thr Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 <210> 13 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(357) <400> 13 cag gtt cag ctg caa gag tct ggc cct ggc ctg gtc aag cct tcc gaa 48 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 aca ctg tcc ctg acc tgc agc gtg acc ggc tac tct atc acc tcc gac 96 Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 tac tac tgg cac tgg atc aga cag cct cca ggc aaa ggc ctg gaa tgg 144 Tyr Tyr Trp His Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 atc gcc tac atc aga tac gac ggc cgg aac gac tac aac ccc agc ctg 192 Ile Ala Tyr Ile Arg Tyr Asp Gly Arg Asn Asp Tyr Asn Pro Ser Leu 50 55 60 aag aac aga gtg acc atc agc cgg gac acc tcc aag aac cag ttc tcc 240 Lys Asn Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 ctg aag ctg tcc tcc gtg acc gct gct gat acc gcc gtg tac tac tgc 288 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 gcc aga gag gac tac ggc tcc tcc tcc ttt gat tac tgg ggc cag ggc 336 Ala Arg Glu Asp Tyr Gly Ser Ser Ser Phe Asp Tyr Trp Gly Gln Gly 100 105 110 acc ctg gtc acc gtt agt tct 357 Thr Leu Val Thr Val Ser Ser 115 <210> 14 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 14 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Tyr Trp His Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Ala Tyr Ile Arg Tyr Asp Gly Arg Asn Asp Tyr Asn Pro Ser Leu 50 55 60 Lys Asn Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Gly Ser Ser Ser Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 15 <211> 324 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(324) <400> 15 gac atc cag atg acc cag tct cca tcc tct ctg tcc gcc tct gtg ggc 48 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 gac aga gtg acc atc acc tgt cgg gcc tcc gag aac atc tac tcc aac 96 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Asn 20 25 30 ctg gcc tgg tat cag cag aag cct ggc aag gct cct aag ctg ctg gtg 144 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 tac gcc gct acc aat ctg gct gat ggc gtg ccc tct aga ttc tcc ggc 192 Tyr Ala Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 tct ggc tct ggc acc gac tat acc ctg aca atc tcc agc ctg cag cct 240 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 gag gac ttc gcc acc tac tac tgc cag cac ttc tgg gac agc cct cca 288 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Phe Trp Asp Ser Pro Pro 85 90 95 acc ttt ggc cag ggc acc aag gtg gaa atc aag cgt 324 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 <210> 16 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 16 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Phe Trp Asp Ser Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 <210> 17 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(357) <400> 17 cag gtt cag ctg caa gag tct ggc cct ggc ctg gtc aag cct tct cag 48 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 acc ctg tct ctg acc tgc agc gtg acc ggc tac tcc atc acc tcc gac 96 Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 tac tac tgg cac tgg atc aga cag cct cca ggc aaa ggc ctg gaa tgg 144 Tyr Tyr Trp His Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 atc gcc tac atc aga tac gac ggc cgg aac gac agc aac ccc agc ctg 192 Ile Ala Tyr Ile Arg Tyr Asp Gly Arg Asn Asp Ser Asn Pro Ser Leu 50 55 60 aag aac aga gtg acc atc agc cgg gac acc tcc aag aac cag ttc tcc 240 Lys Asn Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 ctg aag ctg tcc tcc gtg acc gct gct gat acc gcc gtg tac tac tgc 288 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 gcc aga gag gac tac gcc tcc tcc tcc ttt gat tac tgg ggc cag ggc 336 Ala Arg Glu Asp Tyr Ala Ser Ser Ser Phe Asp Tyr Trp Gly Gln Gly 100 105 110 acc ctg gtc acc gtt agt tct 357 Thr Leu Val Thr Val Ser Ser 115 <210> 18 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 18 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Tyr Trp His Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Ala Tyr Ile Arg Tyr Asp Gly Arg Asn Asp Ser Asn Pro Ser Leu 50 55 60 Lys Asn Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Tyr Ala Ser Ser Ser Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 19 <211> 324 <212> DNA <213> Artificial sequence <220> <223> Mouse antibody / humanized <220> <221> CDS <222> (1)..(324) <400> 19 gac atc cag atg acc cag tct cca tcc tct ctg tcc gcc tct gtg ggc 48 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 gac aga gtg acc atc acc tgt cgg cct tcc gag aac atc tac tcc aac 96 Asp Arg Val Thr Ile Thr Cys Arg Pro Ser Glu Asn Ile Tyr Ser Asn 20 25 30 ctg gcc tgg tat cag cag aag cct ggc aag gct cct aag ctg ctg gtg 144 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 tac gcc gct acc aat ctg gct gat ggc gtg ccc tct aga ttc tcc ggc 192 Tyr Ala Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 tct ggc tct ggc acc gac tat acc ctg aca atc tcc agc ctg cag cct 240 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 gag gat atc gcc acc tac tac tgc cag cac ttc tgg gac agc cct cca 288 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln His Phe Trp Asp Ser Pro Pro 85 90 95 acc ttt ggc cag ggc acc aag ctg gaa atc aag cgt 324 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 <210> 20 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 20 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Pro Ser Glu Asn Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln His Phe Trp Asp Ser Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 <210> 21 <211> 1350 <212> DNA <213> Artificial Sequence <220> <223> Modified Antibody <220> <221> CDS <222> (1)..(1350) <400> 21 cag gtc cag ctg caa gag tct ggc cct gga ctg gtc aag cct tct ggc 48 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly 1 5 10 15 acc ctg tct ctg aca tgt gct gtg tcc ggc ggc tcc atc tcc tcc tct 96 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser 20 25 30 aat tgg tgg tct tgg gtc cga cag cct cct ggc aaa gga ctg gaa tgg 144 Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 atc ggc gag atc tac cac tcc ggc aac acc aac tac aac ccc agc ctg 192 Ile Gly Glu Ile Tyr His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu 50 55 60 aag tcc aga gtg acc gtg tcc gtg gac aag tcc aag aac cag ttc tcc 240 Lys Ser Arg Val Thr Val Ser Val Asp Lys Ser Lys A...

Claims

1. A single-arm anti-human NR1 antibody comprising a heavy-chain variable region and a light-chain variable region, wherein the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence of amino acids 31 to 35 of SEQ ID NO: 10, CDR2 consisting of the amino acid sequence of amino acids 50 to 66 of SEQ ID NO: 10, and CDR3 consisting of the amino acid sequence of amino acids 99 to 111 of SEQ ID NO: 10, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence of amino acids 24 to 34 of SEQ ID NO: 12, CDR2 consisting of the amino acid sequence of amino acids 50 to 56 of SEQ ID NO: 12, and CDR3 consisting of the amino acid sequence of amino acids 89 to 97 of SEQ ID NO:

12.

2. The single-arm anti-human NR1 antibody according to claim 1, which is a single-arm anti-human NR1 antibody comprising a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10 and a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:

12.

3. The single-arm anti-human NR1 antibody according to claim 1, which is a single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy-chain variable region, the heavy-chain variable region comprises CDR1 consisting of the amino acid sequence of amino acids 31 to 35 of SEQ ID NO: 10, CDR2 consisting of the amino acid sequence of amino acids 50 to 66 of SEQ ID NO: 10, and CDR3 consisting of the amino acid sequence of amino acids 99 to 111 of SEQ ID NO: 10, the light chain comprises a light-chain variable region, and the light-chain variable region comprises CDR1 consisting of the amino acid sequence of amino acids 24 to 34 of SEQ ID NO: 12, CDR2 consisting of the amino acid sequence of amino acids 50 to 56 of SEQ ID NO: 12, and CDR3 consisting of the amino acid sequence of amino acids 89 to 97 of SEQ ID NO:

12.

4. The single-arm anti-human NR1 antibody according to claim 3, which is a single-arm anti-human NR1 antibody comprising a heavy chain, a light chain, and an Fc polypeptide, wherein the heavy chain comprises a heavy-chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10, the light chain comprises a light-chain variable region consisting of the amino acid sequence shown in SEQ ID NO:

12.

5. The single-arm anti-human NR1 antibody according to any one of claims 1 to 4, wherein the Fc region comprises LALA mutations L234A and L235A according to the Eu numbering.

6. The single-arm anti-human NR1 antibody according to any one of claims 1 to 4, wherein the Fc region comprises a staphylococcal protein A mutation.

7. The single-arm anti-human NR1 antibody according to any one of claims 1 to 4, wherein the Fc region comprises LALA mutations L234A and L235A and a staphylococcal protein A mutation according to the Eu numbering.

8. The single-chain anti-human NR1 antibody according to any one of claims 1 to 4, wherein the single-chain anti-human NR1 antibody is a human antibody or a humanized antibody.

9. A single-chain anti-human NR1 antibody, comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:

42.

10. A polynucleotide comprising the nucleotide sequence in (a) and / or (b) below: (a) A nucleotide sequence encoding the heavy-chain variable region of the single-chain anti-human NR1 antibody according to claim 1 or claim 2; (b) A nucleotide sequence encoding the light-chain variable region of the single-chain anti-human NR1 antibody according to claim 1 or claim 2.

11. A polynucleotide comprising the nucleotide sequence in (a) and / or (b) below: (a) A nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody according to claim 3 or claim 4; (b) A nucleotide sequence encoding the light chain of the single-chain anti-human NR1 antibody according to claim 3 or claim 4.

12. A polynucleotide comprising the nucleotide sequence in (a) and / or (b) below: (a) A nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30; (b) A nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:

32.

13. An expression vector comprising a polynucleotide comprising the nucleotide sequence in (a) and / or (b) below: (a) A nucleotide sequence encoding the heavy-chain variable region of the single-chain anti-human NR1 antibody according to claim 1 or claim 2; (b) A nucleotide sequence encoding the light-chain variable region of the single-chain anti-human NR1 antibody according to claim 1 or claim 2.

14. An expression vector comprising a polynucleotide comprising the nucleotide sequence in (a) and / or (b) below: (a) A nucleotide sequence encoding the heavy chain of the single-chain anti-human NR1 antibody according to claim 3 or claim 4; (b) A nucleotide sequence encoding the light chain of the single-chain anti-human NR1 antibody according to claim 3 or claim 4.

15. The expression vector according to claim 13 or 14, further comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide.

16. An expression vector comprising a polynucleotide according to (a) and / or (b) below: (a) A polynucleotide comprising a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30; (b) A polynucleotide comprising a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:

32.

17. The expression vector according to claim 16, further comprising a polynucleotide comprising a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO:

42.

18. A host cell transformed with an expression vector selected from the following (a) to (c): (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-arm anti-human NR1 antibody according to claim 1 or claim 2; (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain variable region of a single-arm anti-human NR1 antibody according to claim 1 or claim 2; (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain variable region of a single-arm anti-human NR1 antibody according to claim 1 or claim 2 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain variable region of said antibody.

19. A host cell transformed with an expression vector selected from the following (a) to (c): (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4; (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4; (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of said antibody.

20. A host cell transformed with an expression vector selected from the following (a) to (d): (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of said antibody; (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of said antibody; (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4, an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain of said antibody, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of said antibody; (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain of a single-arm anti-human NR1 antibody according to claim 3 or claim 4, a polynucleotide containing a nucleotide sequence encoding a light chain of said antibody, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide of said antibody.

21. A host cell transformed with an expression vector selected from the following (a) to (j): (a) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30; (b) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32; (c) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32; (d) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32; (e) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42; (f) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42; (g) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42; (h) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32 and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42; (i) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30, a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO: 32, and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42; (j) An expression vector comprising a polynucleotide containing a nucleotide sequence encoding a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 30 and a polynucleotide containing a nucleotide sequence encoding an Fc polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 42, and an expression vector comprising a polynucleotide containing a nucleotide sequence encoding a light chain consisting of the amino acid sequence shown in SEQ ID NO:

32.

22. A method for generating a single-arm anti-human NR1 antibody, which comprises the step of culturing a host cell according to any one of claims 18-21 to express a polypeptide constituting the single-arm anti-human NR1 antibody.

23. A pharmaceutical composition comprising a single-arm anti-human NR1 antibody according to any one of claims 1 to 11 and a pharmaceutically acceptable excipient.

24. The pharmaceutical composition according to claim 23, which is a pharmaceutical composition for treating anti-NMDAR encephalitis.

25. Use of a single-arm anti-human NR1 antibody according to any one of claims 1 to 11 in the preparation of a pharmaceutical composition for treating anti-NMDAR encephalitis.

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