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WO1994018560A1 - Methode de dosage de la proteine humaine tau, materiel prevu a cet effet et methode diagnostique - Google Patents

Methode de dosage de la proteine humaine tau, materiel prevu a cet effet et methode diagnostique Download PDF

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Publication number
WO1994018560A1
WO1994018560A1 PCT/JP1994/000196 JP9400196W WO9418560A1 WO 1994018560 A1 WO1994018560 A1 WO 1994018560A1 JP 9400196 W JP9400196 W JP 9400196W WO 9418560 A1 WO9418560 A1 WO 9418560A1
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Prior art keywords
antibody
human
tau protein
human tau
antibodies
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PCT/JP1994/000196
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English (en)
Japanese (ja)
Inventor
Kenji Hosoda
Hiroshi Eguchi
Tadakatsu Nakamoto
Shinji Kobayashi
Takaharu Kubota
Hiroshi Mori
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Teijin Ltd
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Teijin Ltd
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Priority to AU60104/94A priority Critical patent/AU6010494A/en
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G01N33/6896—Neurological disorders, e.g. Alzheimer's disease

Definitions

  • the present invention relates to a method for measuring tau protein in human cerebrospinal fluid, a kit therefor, and uses related thereto. More specifically, an immunological method that can accurately measure tau protein in human cerebrospinal fluid by distinguishing between persons with central nervous cell disorders and healthy persons who do not, and a kit therefor And related uses.
  • Tau protein is a component of the paired helical filament (PHF), and its change in neurofibrils leads to the death of nerve cells, so it is known that it may be useful in diagnosing Alzheimer's disease.
  • PHF paired helical filament
  • its measurement has mainly been examined immunohistologically in autopsy samples.
  • An antibody may be used for the measurement.
  • an antibody to measure tau protein for example, cerebrospinal fluid is treated with ammonium sulfate, and the tau protein in the crude fraction is blotted using an antibody to confirm its presence.
  • a method has been proposed [Peter Davis et al., Annals of Neurology Vol.22, (4), P.521 (1987)].
  • the antibody (A1z-50) used in this proposal is a crude brain homogenate from a patient with Alzheimer's disease that contains an abnormal phosphorylated tau protein (A-68).
  • the antibody is obtained as an immunogen, and the detection method using this is a complicated method using Western plotting.
  • U.S. Pat. No. 5,134,062 describes the removal of living fibroblasts from humans, culturing the cells in a medium, and indicating the metabolites of Alzheimer's disease from the cells.
  • a method for screening Alzheimer's disease by immunological detection of a substance is described.
  • This US patent shows skin cells as fibroblasts, and PHF (paired helical filament) and tau protein as indicators of metabolites of Alheimer's disease. This method requires removing and culturing the skin cells, and requires complicated means and a long time to obtain a predetermined amount of the indicator substance.
  • the U.S. patent does not describe any specific examples showing the amount of PHF or protein involved.
  • Canadian Patent No. 1,302,250 describes a method of measuring PHF or abnormal phosphorylated protein in cerebrospinal fluid to be useful for treating Alzheimer's disease. This method uses a monoclonal antibody that reacts with PHF.
  • a first object of the present invention is to provide a method for immunologically measuring soluble human tau protein present in a body fluid from a living body, particularly cerebrospinal fluid, and a kit therefor.
  • a second object of the present invention is to provide a method for early and easy measurement of soluble human tau protein in cerebrospinal fluid and a kit therefor.
  • a third object of the present invention is to provide a patient with central nervous
  • An object of the present invention is to provide an immunological method and a kit for accurately measuring soluble human tau protein in cerebrospinal fluid by distinguishing it from that of a healthy human.
  • Another object of the present invention is to provide an immunological method capable of accurately measuring soluble human tau protein therein using a small amount of cerebrospinal fluid.
  • the object of the present invention is to provide a method for immunologically measuring human tau protein in human cerebrospinal fluid, which comprises an insoluble carrier-immobilized antibody and a labeled antibody. And that all antibodies have a recognition site in the amino acid sequence region of positions 251 to 441 of the human tau protein. was found.
  • the object of the present invention is:
  • a kit for immunologically measuring soluble human tau protein in human cerebrospinal fluid wherein the antibodies (a-1) and (a-2) are all the second human tau protein. It has been found that this can be achieved by a kit characterized by having a recognition site in the amino acid sequence region from the 51st to the 44th amino acid.
  • the object of the present invention is as follows.
  • the measurement method of the present invention and the kit therefor are characterized in that both the immobilized antibody and the labeled antibody have the amino acid sequence of human tau protein from the 25th position (Pro.) To the 25th position. It is characterized in that an antibody having a recognition site in the region up to the 1st (Leu.) Is used.
  • the measurement results of the present invention can be used for examination, research, or diagnosis from a clinical or pathological viewpoint of a person having or suspected of having a disorder in central nervous cells.
  • a central nervous cell disorder is a disease or condition caused by any disorder of the central nervous cells, such as a disease or condition caused by cerebrovascular disorder, Alzheimer's disease and carbon monoxide poisoning.
  • the present invention can be expected to be used for examination, research, or diagnosis of Alzheimer's disease or a person suspected of having Alzheimer's disease. All of the above-mentioned diseases based on disorders of central nervous cells are caused by soluble human tau in cerebrospinal fluid. The protein concentration shows a higher value than that of a healthy person.
  • the two antibodies, the immobilized antibody and the labeled antibody are both intermediate regions that are the 251st to the 441st region in the amino acid sequence of 441 of the human tau protein. It has a recognition site in the amino acid sequence region at the C-terminus from the beginning, and it is not clear why highly sensitive and soluble human protein can be measured with it.
  • the present inventors have prepared several monoclonal and polyclonal antibodies specifically recognizing human tau protein and used them for immunoassay of soluble human tau protein in cerebrospinal fluid. Tried. For example, several monoclonal antibodies having a recognition site in the region N-terminal to the amino acid sequence of the amino acid sequence of the human tau protein were obtained. Immunological measurements were performed using a combination of several monoclonal and polyclonal antibodies. A calibration curve could be prepared with such a combination of antibodies, but when cerebrospinal fluid was actually measured as a test sample, the sensitivity was insufficient and stable measurement results could not be obtained. Alternatively, nothing could be used for diagnosis.
  • the antibody used in the present invention may be a monoclonal antibody as long as it has a recognition site in the amino acid sequence of the amino acid sequence of the human tau protein in the 25th to 441st regions.
  • it may be a polyclonal antibody.
  • the combination may be any, but it is preferable to use a polyclonal antibody for both the immobilized antibody and the labeled antibody.
  • the antibody of the present invention may be a polyclonal antibody or a monoclonal antibody as described above.
  • Freund Complete Adjuvant or A1 (OH) 3 is used as an adjuvant. (Haraum) and purified human tau protein Can be obtained as an antiserum by immunizing animals by the method
  • hybridomas prepared by the cell fusion method of Keller and Milstein [G. Koeller and Milstein, Nature (Londa), 256, 495-497 (1975)] are cultured and It is prepared by separating the antibody reacting with the human tau protein from the culture solution.
  • Cells used for cell fusion include P3U1 and the like.
  • the human tau protein used as the immunogen is preferably one that is soluble in Trissalline having a pH of 6.5 to 8.5 (neutral) and purified from human brain.
  • the antibodies obtained by immunizing the thus obtained human tau protein those that recognize the protein [2 51-441] of the amino acid sequence of the human tau protein are selected and used. .
  • the immunoassay can specifically and sensitively measure the human protein in cerebrospinal fluid without modification.
  • immunoassays include enzyme immunoassay (EIA), radioimmunoassay (RIA), and fluorescence immunoassay.
  • EIAs include, for example, “oxygen immunoassay” A publicly known method described in Eiji Ishikawa et al., 2nd edition, Medical Shoin 1982) can be used.
  • EIA may be any of a method called a sandwich method and a competitive method.
  • EIA by the sandwich method, quantification is performed using an antibody against the human tau protein of the present invention, for example, according to the following procedure. That is, one of the two antibodies (the first antibody) is immobilized on a suitable insoluble carrier (for example, a plastic container) (hereinafter, this is referred to as “immobilized antibody”). Next, the surface of the insoluble carrier is coated with a suitable substance (for example, bovine serum albumin) to avoid non-specific binding between the insoluble carrier and the reagent or sample to be measured. Like this The insoluble carrier on which the obtained first antibody is immobilized is brought into contact with the sample for a certain period of time and at a temperature for reaction.
  • a suitable insoluble carrier for example, a plastic container
  • a suitable substance for example, bovine serum albumin
  • the solidified antibody (primary antibody) and the human protein in the sample bind.
  • the insoluble carrier is washed with an appropriate washing solution, and then contacted with a solution (eg, an aqueous solution) of the other antibody (second antibody) against the human tau protein labeled with an appropriate labeling substance (eg, an enzyme) for a certain period of time and at a temperature.
  • the second antibody reacts with the human tau protein bound to the immobilized antibody. This is washed with an appropriate washing solution, and then the amount of the labeling substance labeled on the second antibody which is bound to the immobilized antibody on the insoluble carrier via the tau protein is measured.
  • the above sandwich method can also be carried out by simultaneously mixing an immobilized antibody, a labeled antibody and a sample containing a tau protein, and simultaneously bringing the three into contact with each other for a certain period of time and at a temperature to cause a reaction.
  • both the first antibody and the second antibody may be monoclonal antibodies or polyclonal antibodies, or one may be a monoclonal antibody and the other may be a polyclonal antibody. It can also be used as a monoclonal antibody. However, as described above, it is preferable in terms of sensitivity that both the first antibody and the second antibody are polyclonal antibodies.
  • a competition method for example, a fixed amount of a labeled antibody is allowed to react competitively with an antigen immobilized on a carrier and an antigen to be measured to form a carrier-immobilized antigen-labeled antibody complex, and a washing operation is performed. Then, a method of measuring the amount of the labeled substance of the labeled antibody bound to the carrier, or immobilizing the antibody on the carrier, and allowing the labeled antigen to react with the antigen to be measured in a competitive manner, the carrier-immobilized antibody After the washing operation, a labeled antigen complex is formed, and then the amount of the labeled substance of the labeled antigen bound to the carrier is measured.
  • the antibody may be a full antibody, also reducing the F (ab ') 2 F ( ab') 2 obtained by digesting the antibody with Bae trypsin
  • the Fab 'or antibody Antibody fragments that can bind to an antigen, such as Fab obtained by digestion with papine, can be used as antibodies, and these can be labeled and used as labeled antibodies.
  • the insoluble carrier used in the method for measuring a protein according to the present invention may be any one which is usually used for immunological measurement, for example, polystyrene, polyethylene, polypropylene, polyester, polyacrylonitrile, fluororesin, Examples thereof include polymers such as cross-linked dextran, agarose, and polysaccharide, as well as paper, glass, metal, and combinations thereof.
  • the shape of the insoluble carrier can be various shapes such as tray shape, spherical shape, fibrous shape, rod shape, disk shape, disk shape, container shape, cell, test tube and the like.
  • Enzymes, fluorescent substances, luminescent substances, radioactive substances, and the like can be used as the labeling substance of the labeled antibody.
  • enzymes include peroxidase, alkaline phosphatase, / 3-D-galactosidase, fluorescent substances such as fluorescein isothiosinate and phycopyriprotein, and luminescent substances such as isolucinol and luciferase. lucigenin and the like., and then as the radioactive material may be used, such as 1 2 5 I, 1 3 1 K 1 4 C, 3 H, these are not limited to those illustrated, used to immunoassays Anything that can be done may be used.
  • the labeling agent is an enzyme
  • a substrate and, if necessary, a color former are used to measure its activity.
  • peroxidase hydrogen peroxide is used as a substrate
  • 2,2′-azinodi- (3-ethylbenzthiazoline sulfonic acid) ammonium salt is used as a coloring agent.
  • ABTS 2,2′-azinodi- (3-ethylbenzthiazoline sulfonic acid) ammonium salt
  • ABTS 2,2′-azinodi- (3-ethylbenzthiazoline sulfonic acid) ammonium salt is used as a coloring agent.
  • ABTS 2,2′-azinodi- (3-ethylbenzthiazoline sulfonic acid) ammonium salt
  • 5-aminosalicylic acid 5-aminosalicylic acid
  • 0-phenylenediamine 41-aminoantivirin or 3,3 ', 5,5'-tetramethylbenzidine,
  • the lysing agent (b) may be any of those usually used for immunological assays, and includes, for example, phosphate buffer, Tris-HCl buffer, acetate buffer, and the like. However, those having a pH in the range of 6.0 to 8.0 are shown as preferable examples.
  • a detergent generally used for immunological measurement is also used as it is. Examples include physiological saline, phosphate buffer, Tris-HCl buffer, and mixtures thereof.
  • Non-ionic surfactants such as Triton XI 00, Tween 20 or Brig 35, and ionic surfactants such as sodium dodecyl sulfate may be added to these detergents. Good.
  • Example 1 (Acquisition of anti-human tau protein antibody)
  • the frozen human brain was dissolved in 4 volumes of TS Buffer (2 OmM Tris, 150 mM NaC; pH 7.6).
  • the lysate was centrifuged (10 K rp.mX for 50 minutes), and the supernatant was fractionated using ammonium sulfate (33-50% ammonium sulfate).
  • the resulting precipitate was dissolved in TBS Buffer (50 mM Tris, 500 mM NaCl; pH 7.6; 16 volumes of brain weight), homogenized uniformly, and boiled for 10 minutes. Then, the mixture was centrifuged (10 kr.p.mX10 minutes), and the supernatant was fractionated using Sephadex G-25 (equilibrium solution: 0.1 M MES: pH 6.5).
  • the salt was eluted by changing the salt concentration using phosphocellulose, and concentrated again using saturated ammonium sulfate. Shrunk.
  • the obtained precipitate was suspended in 1 m of TS Buffer, dissolved, filtered through a Millipore filter HV (0.45 / zm), and then subjected to HPLC (column: J.T. Bakerbond C 4 4.6 ⁇ 2). 5 Omm), the second peak eluted was collected. This is the purified human tau protein.
  • the human tau protein purified in (1) above was mixed with Freund's complete adj uvant to prepare WZO / W emulsion. Rabbits were immunized three times, every two months. After three immunizations, whole blood was collected 7 days later. Next, the antiserum was applied to a protein A-Sepharose column equilibrated with 0.1 M phosphate buffer (pH 8.0), washed, and washed with 0.1 M quencher with pH 3.0. The acid Na was allowed to flow, and the anti-human tau protein antibody (IgG) was eluted from the column to obtain a purified antibody (IgG).
  • Example 2 Determination of recognition site of anti-human tau protein antibody
  • Example 1 It is known that human and human tau proteins have highly similar primary structures and also have cross-reactivity in quarantine. Thus, the recognition site of the anti-human tau protein antibody obtained in Example 1 was determined as follows using a large amount of easily prepared dicitau protein.
  • ⁇ sitau protein was cleaved by CNBr (Baker bond C4, column) in the presence of formic acid.
  • the reaction was separated using reverse phase HPLC (acetonitrile 0 ⁇ 80% gradient in 40 min, flow rate lm / min) to give a fraction of 80.
  • Figure 1 shows the reversed phase HPLC pattern.
  • the fractions No.34 and No.44 showed high reactivity. That is, it was clarified that the peptides contained in these fractions specifically bound to the anti-human tau protein antibody obtained in Example 1.
  • the N-terminus of the fraction N 0.34 peak is Pro-Asp-Leu-Lys, and the N-terminus of the No.44 peak may also be Pro-Asp-Leu-Lys. found.
  • Example 3 Measurement of human tau protein by sandwich method EIA using anti-human tau protein antibody
  • Example 1 After thoroughly washing the polystyrene beads (diameter 6 mm), the antibody obtained in Example 1 was allowed to stand in a PBS solution having a concentration of 20 gZm at a temperature of 4 ° C for 24 hours, and then washed with PBS. In a 1% bovine serum albumin (BSA) solution in PBS at a temperature of 4 ° C. for one day and night, post-coating treatment was performed to obtain anti-human tau protein antibody-immobilized beads.
  • BSA bovine serum albumin
  • O.IMEDTA160 a and 1.6 M £ of 1 M hydroxylamine were added and reacted at 37 ° C for 4 minutes. Thereafter, the reaction solution was placed in a collodion bag, and dialyzed at 4 ° C. for 3 days using a solution containing 0.1 M PB (pH 6.0) and 5 mM EDTA to obtain a thiolated HRP.
  • Example 3 Using the method of Example 3 and the calibration curve obtained in Example 3, the concentration of tau protein in the cerebrospinal fluid of 9 patients with Alhaima's disease was measured. As a control, cerebrospinal fluid of 14 healthy subjects was measured. The results are shown in Figure 3. As shown in Fig. 3, a significantly high level of tau protein in cerebrospinal fluid of Alzheimer's disease patients is observed. (P ⁇ 0.001)
  • Alzheimer's disease can be diagnosed in the case of a human protein concentration of 0.5 ngZm or more.
  • Example 5 Measurement of tau protein in patients with cerebrovascular disorder and carbon monoxide poisoning
  • Example 1 Genetics of monoclonal antibody against human tau protein and determination of recognition site
  • Monoclonal antibodies against human tau protein were prepared according to the method of Keraichi and Milstein (Koeller & Milstein). That is, the mouse was immunized by mixing the human tau protein Songhead and Freund's complete adjvant. After four immunizations, fusion was performed three days later. After confirming the appearance of hybridomas, antibody-producing hybridomas were screened using solid-phase tau protein (fixed at 2 g nom).
  • the 8 clones established in Reference Example 1 were cultured, and Lonal antibodies were purified. All of the eight monoclonal antibodies were each used as a 10 ⁇ g / mi solution, and each solution was immersed in a polystyrene ball to immobilize the monoclonal antibodies. A sandwich measurement system using each of the immobilized monoclonal antibodies and the HRP-labeled anti-human tau protein antibody of Example 3 was constructed. Only two of the eight monoclonal antibodies were able to obtain good calibration curves.
  • A, B and C in FIG. 6 show the results of the following measurement systems, respectively.
  • Ala Asp Gly Lys Thr Lys lie A 1 a Thr Pro Arg G 1 y Ala ⁇ 1 a Pro Pro 145 150 155 160
  • Gly Lys Val G 1 n lie lie Asn then ys Lys then eu As then eu Ser Asn Va 1 Gin
  • Lys Lys lie G ⁇ u Thr His or ys Leu Thr Phe Arg G 1 u Asn Ala or ys A 1 a
  • FIG. 1 shows the HPLC pattern of the CN Br degradation product of ⁇ sitau protein.
  • FIG. 2 is a calibration curve obtained in Example 3 (3) Sandwich EIA measurement system.
  • FIG. 3 shows the results of measurement of tau protein concentration in cerebrospinal fluid of Alzheimer's disease patients and healthy subjects performed in Example 4.
  • FIG. 4 shows the results of measurement of tau protein in cerebrospinal fluid of a patient suffering from cerebrovascular disorder and carbon monoxide poisoning in Example 5.
  • FIG. 5 shows respective calibration curves obtained in Reference Example 2 using three types of Sandwich measuring systems.
  • FIG. 6 shows the results of measurement of tau protein in cerebrospinal fluid of healthy individuals and patients with Alzheimer's disease obtained in Example 6 by using three types of sandwich measurement systems.
  • soluble human tau protein in cerebrospinal fluid can be measured simply and with high sensitivity.
  • the measurement can be performed on the extracted human cerebrospinal fluid as it is without any processing, and it is measured as a distinct value between a person with central nervous cell disorder and a healthy person
  • a small amount of cerebrospinal fluid as a test sample is sufficient.
  • the present invention can be effectively used for examination, research, or diagnosis of patients having disorders of central nervous cells.

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Abstract

Méthode de dosage immunologique d'une protéine humaine tau présente dans un fluide cérébrospinal humain, consistant à utiliser un anticorps immobilisé sur un support insoluble et un anticorps marqué, chaque anticorps ayant un site de reconnaissance dans la zone de séquences d'aminoacides comprise entre le 251ème et le 441ème reste d'aminoacides d'une protéine humaine tau; et matériel de dosage immunologique prévu à cet effet. Grâce à cette méthode et à ce matériel, on peut déterminer une concentration de la protéine tau d'un homme normal et celle d'un patient présentant une cytopathie du système nerveux central, concentration qui doit être spécifiée avec une grande précision. Les résultats du dosage peuvent servir pour un test, une étude ou un diagnostic concernant des patients atteints d'une cytopathie du système nerveux central.
PCT/JP1994/000196 1993-02-12 1994-02-10 Methode de dosage de la proteine humaine tau, materiel prevu a cet effet et methode diagnostique Ceased WO1994018560A1 (fr)

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JP4613393A JPH06239899A (ja) 1993-02-12 1993-02-12 ヒトタウ蛋白に対する抗体、並びに該抗体を利用する体液中のヒトタウ蛋白の測定方法
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001055725A3 (fr) * 2000-01-24 2001-12-20 Innogenetics Nv Diagnostic des tauopathies
US7446180B2 (en) 2001-02-02 2008-11-04 Axon Neuroscience Forschungs-Und Entwicklungs Gmbh Conformationally abnormal forms of tau proteins and specific antibodies thereto
EP1995255A1 (fr) 2002-07-12 2008-11-26 Axon Neuroscience Forschungs- und Entwicklungs Gmbh Protéines tau tronquées
US9834596B2 (en) 2012-07-03 2017-12-05 Washington University Antibodies to tau
US9957317B2 (en) 2014-06-27 2018-05-01 C2N Diagnostics, Llc Humanized anti-tau antibodies
CN109580955A (zh) * 2018-11-21 2019-04-05 北京利德曼生化股份有限公司 用于检测Tau蛋白(TAU)的磁微粒分离化学发光免疫测定法
US10251952B2 (en) 2014-06-26 2019-04-09 Hoffmann-La Roche Inc. Humanized anti-tau(pS422) antibody brain shuttles and use thereof
US10465000B2 (en) 2013-12-20 2019-11-05 Hoffmann-La Roche Inc. Humanized anti-Tau(pS422) antibodies and methods of use
US10822402B2 (en) 2015-06-24 2020-11-03 Hoffmann-La Roche Inc. Humanized anti-tau(pS422) antibodies and methods of use
JP2024178283A (ja) * 2018-11-22 2024-12-24 富士レビオ株式会社 抗体コンジュゲート

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* Cited by examiner, † Cited by third party
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JP4624559B2 (ja) * 1998-09-08 2011-02-02 イノジェネティックス・ナムローゼ・フェンノートシャップ 初期ｃｎｓダメージのマーカーとしてのタウ
JP2013032295A (ja) * 2009-12-04 2013-02-14 Kaneka Corp タウ蛋白の吸着材および吸着除去システム

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5716355A (en) * 1980-04-25 1982-01-27 Hoffmann La Roche Immunological method
EP0155224A2 (fr) * 1984-03-15 1985-09-18 CHROMAGENICS, Inc. Complexe en phase solide porteur d'une fonction sensible à un chromogène pour la détection d'un anticorp, d'un antigène, d'un récepteur ou d'un ligand
US5134062A (en) * 1988-03-22 1992-07-28 Cornell Research Foundation, Inc. Diagnosis of neuronal disorders and screening potential therapeutic agents therefor
FR2678638A1 (fr) * 1991-07-03 1993-01-08 Inst Nat Sante Rech Med Anticorps polyclonaux et monoclonaux contre la proteine tau pour le diagnostic de la maladie d'alzheimer.
WO1993008302A1 (fr) * 1991-10-25 1993-04-29 Itt Automotive Europe Gmbh Anticorps monoclonaux diriges contre la proteine tau associee aux microtubules
WO1993011231A1 (fr) * 1991-12-06 1993-06-10 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Outils de diagnostic et de traitement de la maladie d'alzheimer

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5716355A (en) * 1980-04-25 1982-01-27 Hoffmann La Roche Immunological method
EP0155224A2 (fr) * 1984-03-15 1985-09-18 CHROMAGENICS, Inc. Complexe en phase solide porteur d'une fonction sensible à un chromogène pour la détection d'un anticorp, d'un antigène, d'un récepteur ou d'un ligand
US5134062A (en) * 1988-03-22 1992-07-28 Cornell Research Foundation, Inc. Diagnosis of neuronal disorders and screening potential therapeutic agents therefor
FR2678638A1 (fr) * 1991-07-03 1993-01-08 Inst Nat Sante Rech Med Anticorps polyclonaux et monoclonaux contre la proteine tau pour le diagnostic de la maladie d'alzheimer.
WO1993008302A1 (fr) * 1991-10-25 1993-04-29 Itt Automotive Europe Gmbh Anticorps monoclonaux diriges contre la proteine tau associee aux microtubules
WO1993011231A1 (fr) * 1991-12-06 1993-06-10 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Outils de diagnostic et de traitement de la maladie d'alzheimer

Non-Patent Citations (12)

* Cited by examiner, † Cited by third party
Title
ACTA NEUROPATHOLOGICA, Vol. 80, No. 2, (1990), DELACOURTE A. et al., "Pathological Proteins Tau 64 and 69 are Specifically Expressed in the Somatodendritic Domain of the Degenerating Cortical Neurons During Alzheimer's Disease Demonstration with a Panel of Antigodies Against Tau Proteins", p. 111-117. *
ALZHEIMER DISEASE AND ASSOCIATED DISORDERS, Vol. 6, No. 4, (1992), CAPUTO C.B. et al., "Immunoreactivity of an Antibody to a Beta-A4 Sequence with Alzheimer Paired Helical Filaments and Tau Protein", p. 225-235. *
BIOCHEMICAL JOURNAL, Vol. 273, No. 1, (1991), BRION J-P et al., "Tau in Alzheimer Neurofibrillary Tangles Amino and Carboxyl-Terminal Regions are Differentially Associated with Paired Helical Filaments and the Location of a Putative Abnormal Phosphorylation Site", p. 127-134. *
BRAIN RESEARCH, Vol. 629, No. 1, (1993), KENESSEY A. et al., "The Extent of Phosphorylation of Fetal Tau is Comparable that of PHF-Tau from Alzheimer Paired Helical Filaments", p. 40-46. *
JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. 268, No. 13, (1993), POULTER L. et al., "Locations and Immunoreactivities of Phosphorylation Sites on Bovine and Porcine Tau Proteins and a PHF-Tau Fragment", p. 9636-9644. *
JOURNAL OF IMMUNOLOGICAL METHODS, Vol. 134, No. 2, (1990), HARRINGTON C.R. et al., "Competitive ELISA for the Measurement of Tau Protein in Alzheimer's Disease", p. 261-272. *
JOURNAL OF NEUROSCIENCE, Vol. 10, No. 10, (1990), UEDA K et al., "Alz-50 Recognizes a Phosphorylated Epitope of Tau Protein", p. 3295-3304. *
NEUROSCIENCE, Vol. 37, No. 1, (1990), NIETO A. et al., "Characterization of Tau Protein Present in Microtubules and Paired Helical Filaments of Alzheimer's Disease Patient's Brain", p. 163-170. *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol. 86, No. 14, (1989), IQBAL K. et al., "Identification and Paired Helical Filaments of Alzheimer Disease", p. 5646-5650. *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol. 88, No. 13, (1991), HARRINGTON C.R. et al., "Measurement of Distinct Immunochemical Presentations of Tau Proteins in Alzheimer Disease", p. 5842-5846. *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol. 88, No. 13, (1991), NOVAK M. et al., "Difference Between the Tau Protein of Alzheimer Paired Helical Filament Core and Normal Tau Revealed by Epitope Analysis of Monoclonal Antibodies 423 and 7. 51", p. 5837-5841. *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol. 89, No. 12, (1992), LICHTENBERG-KRAAG B. et al., "Phosphorylation-Dependent Epitopes of Neurofilament Antibodies on Tau Protein and Relationship with Alzheimer Tau", p. 5384-5388. *

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JP2003521499A (ja) * 2000-01-24 2003-07-15 インノジェネティクス・エヌ・ブイ タウオパチーの診断
US6680173B2 (en) 2000-01-24 2004-01-20 Innogenetics N.V. Diagnosis of tauopathies
US7387879B2 (en) 2000-01-24 2008-06-17 Innogenetics N.V. Diagnosis of tauopathies
US7446180B2 (en) 2001-02-02 2008-11-04 Axon Neuroscience Forschungs-Und Entwicklungs Gmbh Conformationally abnormal forms of tau proteins and specific antibodies thereto
US8288608B2 (en) 2002-07-12 2012-10-16 Axon Neuroscience Se Transgenic animal expressing alzheimer's tau protein
EP1995255A1 (fr) 2002-07-12 2008-11-26 Axon Neuroscience Forschungs- und Entwicklungs Gmbh Protéines tau tronquées
US9485972B2 (en) 2002-07-12 2016-11-08 Axon Neuroscience Se Truncated tau proteins
US9834596B2 (en) 2012-07-03 2017-12-05 Washington University Antibodies to tau
US10465000B2 (en) 2013-12-20 2019-11-05 Hoffmann-La Roche Inc. Humanized anti-Tau(pS422) antibodies and methods of use
US10251952B2 (en) 2014-06-26 2019-04-09 Hoffmann-La Roche Inc. Humanized anti-tau(pS422) antibody brain shuttles and use thereof
US9957317B2 (en) 2014-06-27 2018-05-01 C2N Diagnostics, Llc Humanized anti-tau antibodies
US10822402B2 (en) 2015-06-24 2020-11-03 Hoffmann-La Roche Inc. Humanized anti-tau(pS422) antibodies and methods of use
CN109580955A (zh) * 2018-11-21 2019-04-05 北京利德曼生化股份有限公司 用于检测Tau蛋白(TAU)的磁微粒分离化学发光免疫测定法
JP2024178283A (ja) * 2018-11-22 2024-12-24 富士レビオ株式会社 抗体コンジュゲート

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