WO2002050121A1 - Nouvel anticorps - Google Patents

Nouvel anticorps Download PDF

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Publication number
WO2002050121A1
WO2002050121A1 PCT/JP2001/010919 JP0110919W WO0250121A1 WO 2002050121 A1 WO2002050121 A1 WO 2002050121A1 JP 0110919 W JP0110919 W JP 0110919W WO 0250121 A1 WO0250121 A1 WO 0250121A1
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Prior art keywords
antibody
synuclein
phosphorylated
seq
peptide
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PCT/JP2001/010919
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English (en)
Japanese (ja)
Inventor
Takeshi Iwatsubo
Hideo Fujiwara
Masato Hasegawa
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Taisho Pharmaceutical Co.,Ltd.
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Priority to JP2002552014A priority Critical patent/JP4055579B2/ja
Priority to AU2002221132A priority patent/AU2002221132A1/en
Publication of WO2002050121A1 publication Critical patent/WO2002050121A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans

Definitions

  • the present invention relates to an antibody that specifically binds to ⁇ -synuclein aggregates such as Levibody, a Levibody containing the antibody, an agent for detecting a synucleinopathy (synucleinopathy) lesion, and a Lewy body. And a method for detecting synucleinopathic (synucleinopathy) lesions.
  • Degenerative neurons such as Parkinson's disease (PD) and Lewy body dementia (DLB) accumulate «-synuclein in the form of Lewy bodies (LB) [Am. J. Pathol, 152 879 (1998)].
  • -Synuclein is also known to accumulate in glial cells of multiple system atrophy, a type of spinocerebellar degeneration, LB in Hallervorden-Spatz disease, degenerative neurites, etc. in addition to ⁇ D and DLB. It is collectively called one (synucleinopathy).
  • the discovery of mutations in a-synuclein in familial PD families [Science 276: 2045 (1997)] has highlighted the role of ⁇ -synuclein in PD and in synucleinopathy.
  • Ser of nuclein can be phosphorylated by G protein-coupled receptor kinase (GRK), casein kinase (CK1, CK2), etc. [J Biol Chem 275, 26515 (2000), J Biol Chem 275, 390] (2000)], but under physiological conditions There are no reports that the above-mentioned Lewy bodies are phosphorylated under pathological conditions such as PD or other synucleinopathies. Disclosure of the invention
  • the present inventors have found that a part of the constituent amino acids is phosphorylated in hynuclein, which forms an aggregate such as Levi body. Furthermore, they have found that an antibody prepared using the synthetic peptide having the phosphorylated amino acid as an antigen specifically binds to ⁇ -synuclein aggregates, thereby completing the present invention.
  • the present invention is an antibody that specifically binds to a protein in which one or more Ser in SEQ ID NO: 1 is phosphorylated.
  • the present invention is an antibody that specifically binds to a protein in which any one of Ser9, Ser42, Ser87 or Serl29 in SEQ ID NO: 1 is phosphorylated.
  • the present invention is an antibody that specifically binds to a protein in which Serl 29 is phosphorylated in SEQ ID NO: 1.
  • the present invention is an antibody that specifically binds to a peptide in which Ser6 is phosphorylated in SEQ ID NO: 2.
  • the present invention is also an antibody that specifically binds to a protein in which Ser87 is phosphorylated in SEQ ID NO: 1.
  • the present invention is an antibody that specifically binds to a phosphorylated Ser6 peptide in SEQ ID NO: 3.
  • the present invention is a drug for detecting a lesion of synucleinopathy, which comprises any one of the above antibodies.
  • the present invention provides a method for detecting ⁇ -synuclein in which Serl29 or Ser87 is phosphorylated in SEQ ID NO: 1, which comprises using any of the above antibodies.
  • Figure 1 shows the results of Western blot analysis of -synuclein extracted from DLB brain. Represent. The fraction obtained by solubilizing DLB brain with 50 mM Tris buffer (Tris HC1), 1% Triton-X, 1% SarkosyK 8 M urea (Urea) was used for anti-human ⁇ -synuclein antibody LB509. Western blot analysis. C represents normal control human brain, and D represents DLB patient brain. The molecular weight marker position (in kilodaltons) is shown on the left. The ⁇ -synuclein protein band is positive at 15 kilodaltons.
  • Tris buffer Tris buffer
  • Triton-X Triton-X
  • Urea SarkosyK 8 M urea
  • FIG. 2 shows the results of staining of DL L brain tissue with an anti-synuclein antibody and a phosphorylation-specific anti-synuclein antibody.
  • DLB cerebral cortex After fixing DLB cerebral cortex to formalin, slice into 50-micron thickness, and use avidin-biotin complex method with anti-human spleen synuclein antibody LB509 ( ⁇ ) and Ser 129 phosphorylation specific ⁇ -synuclein antibody ( ⁇ ). And stained with diaminobenzidine to develop a brown color.
  • LB shows a positive circle (arrow), and normal -synuclein shows a diffuse, fine-granular, positive staining throughout the neuropil, but a negative neuronal cell body (*) .
  • B in addition to L B (arrow), a short shrunken Lewy neuri te (arrow head) is clearly depicted.
  • FIG. 3 shows the results of Western blot analysis using anti-synuclein specific anti-synuclein antibody extracted from DLB brain.
  • the fraction obtained by solubilizing DLB brain with 50 mM Tris buffer (Tris HC1), 1% Triton-X, 1% SarkosyK 8 M urea (Urea) was used for Serl29 phosphorylation-specific ⁇ -synuclein antibody.
  • C represents normal control human brain
  • D represents DLB patient brain.
  • the position of the molecular weight marker (displayed in mouth Dalton) is shown on the left.
  • a band of phosphorylation ⁇ -synuclein protein is positive at a position of 15 kDa specifically for the urea-soluble fraction.
  • FIG. 4 shows an ELISA study of the specificity of Ser 129 phosphorylated ⁇ -synuclein-specific antibody after affinity purification.
  • the amount of Serl29 phosphorylated peptide shown on the horizontal axis (sequence is as in Example 2) or recombinant full-length human synuclein is fixed to the microwell plate, and reacted with Ser129-phosphorylated human synuclein-specific antibody. Color was developed using TMB Microwell Peroxidase Substrate (Funakoshi).
  • Serl29 phosphorylation ⁇ -synuclein-specific antibody reacts specifically with Serl29 phosphorylated peptide, but does not react with non-phosphorylated recombinant human synuclein.
  • FIG. 5 shows an ELISA study of the specificity. Serl29 phosphorylation in the amount indicated on the horizontal axis (The sequence is as in Example 5.) Alternatively, the total length of the recombinant ⁇ ⁇
  • FIG. 6 shows the results of Western blot analysis of 0! -Synuclein extracted from DLB brain.
  • Western blot was performed using the fraction obtained by solubilizing DLB brain with 50 mM Tris buffer (Tris HC1), 1% Triton-X, 1% SarkosyK 8 M urea (Urea), and SDS. Lot analysis was performed. The position of the molecular weight marker (in kilodaltons) is shown on the left. The band of a-synuclein protein is positive at 15 kilodaltons.
  • FIG. 7 shows the EUSA study of the specificity of Ser87 phosphorylated ⁇ -synuclein-specific antibody after affinity purification.
  • the amount of Ser87 phosphorylated peptide indicated on the horizontal axis (sequence is as in Example 8) or recombinant full-length synuclein is immobilized on a microwell plate, and after reaction with Serl 29 phosphorylated sph-synuclein-specific antibody, TMB Mi crowel The color was developed using l Peroxi dase Substrate (Funakoshi).
  • Ser87 phosphorylated ⁇ -synuclein specific antibody reacts specifically with Ser87 phosphorylated peptide, but does not react with non-phosphorylated recombinant ⁇ -synuclein.
  • a protein in which one or two or more Sers in SEQ ID NO: 1 is phosphorylated refers to a protein in which any of Ser, 9, 42, 87 or 129 in SEQ ID NO: 1 is phosphorylated
  • the phosphorylated Ser may be present at one site or at multiple sites.
  • Serl 29 means Ser which is the 129th amino acid from the N-terminus in the amino acid sequence
  • Ser 6 means Ser which is the 6th amino acid from the N-terminus in the amino acid sequence
  • Ser87 means the amino acid Ser means the amino acid located at the 87th position from the N-terminus in the sequence.
  • the term “specifically binds” means that it binds (or reacts) to human synuclein in which Ser of SEQ ID NO: 1 is phosphorylated, but binds (or reacts) to non-phosphorylated human synuclein. Means no reaction). For example, in SEQ ID NO: 1, it binds to a protein in which Serl29 is phosphorylated, but does not bind to a protein in which Serl29 is not phosphorylated.
  • Ser-phosphorylated peptides or full-length ⁇ -synuclein give a positive reaction by ELISA or Western blot, but do not react with non-phosphorylated Ser or peptide. Means that.
  • an “antibody” is a polyclonal or monoclonal antibody that specifically binds to a fragment containing synuclein and phosphorylated Ser consisting of a full-length molecule, or a partial fragment of these antibodies (eg, papain or It means a fragment (Fab or F (al ') 2 or Fab') obtained by digestion with pepsin, and can be produced according to a known production method as described later.
  • “Synucleinopathic lesion detection drug” refers to the immunohistological study of the brain tissue of synucleinopathy that exhibits LB or other ⁇ -synuclein-positive lesions (eg, degenerative neurites such as GCI and Lewy neurite). Antibodies, reagents or drugs that show a positive reaction or specifically react with insoluble / accumulated human synuclein by Western blot analysis of brain tissue.
  • the antibody according to the present invention can be produced according to the following production method.
  • the DLB brain is fractionated into soluble and insoluble fractions using various surfactants such as Triton-X and Sarkosyl. Further, the insoluble fraction is dissolved in urea, and purified using an anion exchange column, for example, a Q-sepharose column. After digesting the purified product with cyanogen bromide and separating the peptide fragment by HPLC, the Ser phosphorylated target product can be obtained. Also, synthetic peptides containing phosphorylated Ser, for example,
  • Immunization is carried out by administering the antigen peptide solution to a warm-blooded animal by itself or with a carrier or diluent about 2 to 10 times in total.
  • the warm-blooded animals used include, for example, egrets, dogs, guinea pigs, mice and rats. It is preferable that a sample blood be taken at the time of subcutaneous immunization four to six times to measure the antibody titer.
  • the measurement of the antibody titer in the serum can be carried out by fixing the peptide used as the antigen to a 96-well microtiter plate and ELISA.
  • the whole blood can be collected and the antibody can be separated and purified by a commonly used method.
  • the purification method include a gel filtration method and a purification method using an active adsorbent such as protein A. Specificity for phosphorylated ⁇ -synuclein can be improved.
  • Monoclonal antibody-producing cells are prepared by selecting individuals with antibody titers from warm-blooded animals immunized with the antigen, collecting spleen or lymph nodes 2 to 5 days after the final immunization, and producing the antibody contained in them. By fusing the cells with myeloma cells, monoclonal antibody-producing eight hybridoma cells can be prepared. The fusion operation can be performed according to a known method, for example, the method of Kohler et al. (Nature, 256, 495 (1975)). Examples of myeloma cells include PAI and P3U1.
  • fusion promoter examples include polyethylene glycol (PEG) and Sendai virus (HVJ), preferably PEG having a molecular weight of 1,000 to 6,000.
  • PEG polyethylene glycol
  • HVJ Sendai virus
  • Cell fusion can be performed efficiently by adding at a concentration of about 10 to 80% and incubating at 20 to 40 ° C.
  • Selection of the monoclonal antibody can be performed according to a known method. Usually in animal cell culture medium supplemented with HAT (hypoxanthine, aminopterin, thymidine) Done. As a selection and breeding medium, for example, RPMI 1640 medium containing 10 to 20% fetal bovine serum can be used. The cultivation is usually performed under 5% CO 2 at a culture temperature of 20 to 40 for 5 days to 3 weeks.
  • HAT hypoxanthine, aminopterin, thymidine
  • the culture supernatant is collected from the wells in which the hybridoma cells have been cultured, and antibodies that react with the antigen peptide are selected by ELISA (enzyme-linked immunosorbent assay).
  • ELISA enzyme-linked immunosorbent assay
  • the desired hybridoma cells thus obtained are cultured, and a monoclonal antibody can be obtained from the culture supernatant.
  • the eight hybridoma cells can be transformed into, for example, a mouse (
  • Balb / c can be administered intraperitoneally to obtain monoclonal antibodies from the ascites fluid. Purification of the monoclonal antibody can be carried out in the same manner as ordinary separation and purification of a polyclonal antibody. Industrial applicability
  • Example 1 Western blot analysis of ⁇ -synuclein extracted from DLB brain
  • Gray matter was cut out from the cerebral cortex of the DLB brain and the normal brain, and was solubilized stepwise using various surfactants as follows.
  • the cerebral cortex was treated with Tris A solution pH 7.5 [50 mM Tris (Gibco BRL), 1 mM EGTA (Wako Pure Chemical Industries), 0.5 mM PMSF (Boehringer Mannheim),
  • the obtained supernatant is 1000 g sup, and the obtained precipitate is lOOOOg ppt.
  • the lOOOOg sup was centrifuged in a centrifuge at 4 ° (:, 15 minutes, 350,000 X g. The supernatant was used as the Tris-soluble fraction.
  • Triton-X solution Tris A solution, 1% Triton-X 100 (Wako Pure Chemical), 10% sucrose (Kanto Chemical), 0.5 M NaCl (Kanto Chemical)]
  • Triton-X solution Tris A solution, 1% Triton-X 100 (Wako Pure Chemical), 10% sucrose (Kanto Chemical), 0.5 M NaCl (Kanto Chemical)
  • the supernatant was a Triton-X soluble fraction.
  • the resulting precipitate was homogenized in a Sarkosyl solution [50 mM Tris pH7.5, ImM EGTA, 1% Sarkosyl (Wako Pure Chemical Industries)], and centrifuged at 350,000 Xg at 25 ° C for 15 minutes with a centrifuge. .
  • the supernatant was a Sarkosyl soluble fraction.
  • the precipitate thus obtained was again homogenized in Sarkosyl solution, and then centrifuged at 350,000 X g at 25 ° C for 15 minutes in a centrifuge.
  • the Sarkosyl-insoluble precipitate obtained here was homogenized in Tris A solution pH 7.5, and centrifuged at 350,000 X g for 25 to 15 minutes using a centrifuge.
  • the resulting precipitate is sonicated in a urea solution [50 mM Tris pH 7.5, ImM EGTA, 8 M urea (nacalai tesque)] using an ultrasonic crusher (Brans on) and then placed in a 37 ° C water bath. Let stand for 30 minutes. Then, it was centrifuged at 350,000 X g at 25 ° C for 15 minutes. This supernatant is used as the urea-soluble fraction.
  • Tris-soluble fraction, Triton-X-soluble fraction, Sarkosyl-soluble fraction, and urea-soluble fraction thus obtained were electrophoresed using SDS-PAGE (SDS polyacrylamide gel electrophoresis), and immobilon membrane ( MILLIPORE) and the immunoblotting method [Proc. Natl. Acad. Sci. USA, 94] using the monoclonal antibody LB509 [Am. J. Pathol, 152 879 (1998)] that specifically recognizes human ⁇ -synuclein. 2025 (1997)], normal soluble human synuclein was recovered in DLB brain, Tris-soluble fraction of normal brain, and Triton-X soluble fraction.
  • SDS-PAGE SDS polyacrylamide gel electrophoresis
  • MILLIPORE immobilon membrane
  • Normal human synuclein was purified as follows. DL Add ammonium sulfate (Kanto Chemical) to the Tris-soluble fraction obtained from the cerebral cortex of the normal and normal brains to a final concentration of 50%, leave on ice for 30 minutes or more, and centrifuge for 4 minutes. Centrifuge at 350,000 X g for 15 minutes at ° C Released. Remove the supernatant, suspend the precipitate in Tris B solution pH 7.5 [50 mM Tris, ImM EGTA, 1% 2-mercaptoethanol (Kanto Chemical), 0.5 M NaCl], and heat treat [100 ° C, 5 minutes] Then, the supernatant was recovered by centrifugation.
  • Ammonium sulfate Kanto Chemical
  • the fraction with a molecular weight of about 15 kDa which was confirmed to contain ⁇ -synuclein by Western blotting using SDS-PAGE, was subjected to reverse phase HPLC using an Aquapore RP300 column (2.1 ⁇ 30 mm, Applied Biosystems). Was separated into fractions with acetonitrile (Kanto Chemical) concentration of about 60%.
  • ⁇ -synuclein contained in the urea-soluble fraction derived from DLB brain was purified as follows. First, the protein contained in the urea-soluble fraction was adsorbed on a Q-sepharose column (Pharmacia Biotech) and adsorbed stepwise with a urea solution containing 0 M, 0.1 M, 0.2 M, 0.3 M, and 0.5 M NaCl. When the protein was eluted, synuclein eluted in the 0.3 M NaCl fraction. This fraction was fractionated by reverse-phase HPLC using an Aquapore RP300 column. —Synuclein was recovered in a fraction with an acetonitrile concentration of about 60%.
  • the Hishi nuclein contained in the Tris-soluble fraction and urea-soluble fraction purified using HPLC in this manner was dried with a freeze dryer and suspended in a 70% formic acid (Wako Pure Chemical) solution. Peptide bonds were chemically cleaved with 0.2% cyanogen bromide (Nacalai tesque). After the reaction, the solution was diluted about 10-fold, dried with a freeze dryer, and suspended in an 8 M aqueous solution of guanidine hydrochloride (Nacalai Tesque). The peptide fragments thus obtained were separated and fractionated by reversed-phase HPLC using a Superspher Select B column (2.1 ⁇ 125 thigh, Merck).
  • the separated fractions were analyzed with a TOF (time of flight) mass spectrometer (PerSeptive Biosystem) and an amino acid sequence analyzer (Applied Biosystems) [J. Biol. Chem. 267 17047 (1992)].
  • TOF time of flight
  • Applied Biosystems amino acid sequence analyzer
  • a fraction with an acetonitrile concentration of about 31% was added to a fraction with a mass number of 1232 corresponding to the C-terminal part of 117 to 127 of the synuclein, With a mass number of 1515 corresponding to the end portion of 128 to 140 A signal was detected.
  • Example 3 Staining of pathological tissue and Western blot analysis Serl29 phosphorylation—Synuclein-specific antibody (anti-PSerl 29) prepared in Example 2 was used to stain PD and DLB brains.
  • Example 1 fractions obtained by gradually solubilizing the cerebral cortex of DLB brain and normal brain using various surfactants were analyzed by Western blotting using SDS-PAGE.
  • the human synuclein-specific antibody does not react with the normal human synuclein contained in the Tris-soluble fraction and the Triton-X soluble fraction, but specifically with only ⁇ -synuclein contained in the urea-soluble fraction. Reacted ( Figure 3).
  • Facial - it comprises a sequence of 124-134 of synuclein, and peptides containing the Ser 129 phosphorylated, CAYEMPS (P0 3 H 2) was KLH conjugated EEGYQ, and an antigen. 0.5% SDS was added to 100 1 of a 1 mg / ml antigenic peptide physiological saline solution, emulsified with Freund's complete adjuvant, and immunized on the back of a mouse (Balb / c, 6 weeks old).
  • a booster immunization was performed with 500 lmg / ml antigen peptide physiological saline solution, 0.5% SDS, and Freund's incomplete adjuvant emulsified by ultrasonic treatment, and thereafter, booster immunization was performed every week.
  • the spleen was removed, and lymphocytes were removed in RPMI 1640 medium (containing penicillin and streptomycin), and subjected to erythrocyte treatment with 0.17 M ammonium chloride.
  • the extracted lymphocytes were fused with the mouse myeloma-derived myeloma PAI strain by the polyethylene glycol method (PEG4000) to prepare hybridoma cells.
  • the hybridoma cells were suspended in a HAT medium containing one feeder cell, dispensed into a 96-well plate (Greiner), and cultured for 15 days.
  • Example 6 Screening of monoclonal antibody (mAb PSerl29)
  • the culture supernatant was collected from the wells in which the hybridoma cells were cultured, and mAb PSerl29, which reacts with the antigenic peptide, was selected by ELISA (enzyme-linked immunosorbent assay).
  • ELISA enzyme-linked immunosorbent assay
  • mice (Balb / c) to which 0.5 ml of pristane was intraperitoneally administered 7 days and 3 days before, respectively, were intraperitoneally injected with the selected mAb PSerl 29 hybridoma cells, and about 10 days later, ascites was collected.
  • the collected ascites was left at room temperature for 30 minutes, left still at 4 ° C, centrifuged at 15K rpm for 10 minutes, and the supernatant was collected.
  • Example 7 Specificity of Monoclonal Antibody (mAb PSerl29)
  • one-synuclein antibody (ascites) was diluted 1000-fold and allowed to react to develop color, but it did not bind to recombinant ⁇ -synuclein. It reacted specifically and strongly with the antigen peptide (Fig. 5).
  • Facial - comprises 82- 92 array of synuclein, and polypeptides comprising Ser87 phosphorylated [KLH] - CVEGAGS (P0 3 3 ⁇ 4) were synthesized IAAAT (Peptide Institute) was the antigen.
  • ⁇ -synuclein antibody prepared in Example 8 was an antibody specific to Ser87 phosphorylation ⁇ -synuclein.
  • Example 8 After adsorbing 1.25 gell of recombinant ⁇ -synuclein or an antigen peptide (used for antibody production) on the bottom of a 96-well plate (Greiner), the human synuclein antibody prepared in Example 8 was diluted 100-fold. When the reaction was carried out, no binding was observed with the recombinant ⁇ -synuclein, but it was strongly reactive with the antigen peptide.

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Abstract

La présente invention concerne un anticorps qui se lie de manière spécifique à une protéine ayant une séquence d'acides aminés dérivée de la séquence de SEQ ID NO :1 par phosphorylation d'un ou de plusieurs SER; et des médicaments permettant de détecter des lésions de synucléinopathie se caractérisant par le fait qu'ils renferment cet anticorps.
PCT/JP2001/010919 2000-12-13 2001-12-12 Nouvel anticorps WO2002050121A1 (fr)

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JP2002552014A JP4055579B2 (ja) 2000-12-13 2001-12-12 新規抗体
AU2002221132A AU2002221132A1 (en) 2000-12-13 2001-12-12 Novel antibody

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Cited By (6)

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Publication number Priority date Publication date Assignee Title
US20170204173A1 (en) * 2003-05-19 2017-07-20 Prothena Biosciences Limited Antibodies to Alpha Synuclein
JP2018091844A (ja) * 2016-11-29 2018-06-14 国立大学法人 長崎大学 α−シヌクレイン検出方法
JP2021513560A (ja) * 2018-02-12 2021-05-27 ザ スクリップス リサーチ インスティテュートＴｈｅ Ｓｃｒｉｐｐｓ Ｒｅｓｅａｒｃｈ Ｉｎｓｔｉｔｕｔｅ パーキンソン病およびシヌクレイノパチーに関連する方法
US11142570B2 (en) 2017-02-17 2021-10-12 Bristol-Myers Squibb Company Antibodies to alpha-synuclein and uses thereof
US11261242B2 (en) 2017-12-15 2022-03-01 UCB Biopharma SRL Anti-alpha-synuclein antibodies
US11292831B2 (en) 2017-12-15 2022-04-05 UCB Biopharma SRL Anti-alpha synuclein antibodies

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CN111544584A (zh) * 2020-04-16 2020-08-18 首都医科大学 一种单克隆抗体对帕金森病的治疗作用

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JP2000034300A (ja) * 1998-07-17 2000-02-02 Mitsubishi Chemicals Corp 抗リン酸化タウ蛋白質抗体及びそれを用いるアルツハイマー病の検出方法

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WO1999024473A1 (fr) * 1997-11-07 1999-05-20 Aventis Pharma Deutschland Gmbh Anticorps contre une phosphoproteine phosphorylee stimulee par un vasodilatateur (vasp), cellules d'hybridome permettant leur preparation et utilisation desdits anticorps
JP2000034300A (ja) * 1998-07-17 2000-02-02 Mitsubishi Chemicals Corp 抗リン酸化タウ蛋白質抗体及びそれを用いるアルツハイマー病の検出方法

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170204173A1 (en) * 2003-05-19 2017-07-20 Prothena Biosciences Limited Antibodies to Alpha Synuclein
JP2018091844A (ja) * 2016-11-29 2018-06-14 国立大学法人 長崎大学 α−シヌクレイン検出方法
US11142570B2 (en) 2017-02-17 2021-10-12 Bristol-Myers Squibb Company Antibodies to alpha-synuclein and uses thereof
US11827695B2 (en) 2017-02-17 2023-11-28 Bristol-Myers Squibb Company Antibodies to alpha-synuclein and uses thereof
US11261242B2 (en) 2017-12-15 2022-03-01 UCB Biopharma SRL Anti-alpha-synuclein antibodies
US11292831B2 (en) 2017-12-15 2022-04-05 UCB Biopharma SRL Anti-alpha synuclein antibodies
JP2021513560A (ja) * 2018-02-12 2021-05-27 ザ スクリップス リサーチ インスティテュートＴｈｅ Ｓｃｒｉｐｐｓ Ｒｅｓｅａｒｃｈ Ｉｎｓｔｉｔｕｔｅ パーキンソン病およびシヌクレイノパチーに関連する方法

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