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WO1995005395A1 - Procede de diagnostic de la maladie d'alzheimer - Google Patents

Procede de diagnostic de la maladie d'alzheimer Download PDF

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Publication number
WO1995005395A1
WO1995005395A1 PCT/JP1994/001359 JP9401359W WO9505395A1 WO 1995005395 A1 WO1995005395 A1 WO 1995005395A1 JP 9401359 W JP9401359 W JP 9401359W WO 9505395 A1 WO9505395 A1 WO 9505395A1
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WIPO (PCT)
Prior art keywords
protein
disease
alzheimer
antibody
electrophoresis
Prior art date
Application number
PCT/JP1994/001359
Other languages
English (en)
Japanese (ja)
Inventor
Kiminobu Sugaya
Hiroshi Marusawa
Original Assignee
Fujisawa Pharmaceutical Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fujisawa Pharmaceutical Co., Ltd. filed Critical Fujisawa Pharmaceutical Co., Ltd.
Priority to AU74670/94A priority Critical patent/AU7467094A/en
Publication of WO1995005395A1 publication Critical patent/WO1995005395A1/fr

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Classifications

    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4711—Alzheimer's disease; Amyloid plaque core protein

Definitions

  • the present invention relates to a protein useful for studying the cause and treatment of Alzheimer's disease, a therapeutic study, an antibody recognizing the protein and a fragment of the protein, a method for diagnosing Alzheimer's disease using the antibody or the protein, and the like. It relates to the kit used for the diagnosis and is used in the medical field.
  • Alzheimer's disease develops in the elderly and in old age, with progressive dementia as the main symptom, as well as mental symptoms and problematic behaviors associated with dementia: excitement, violence, wandering, hallucinations, delusional insomnia, delirium Pathologically characterized by general brain atrophy, extensive neuronal loss, senile plaques, amyloid protein deposition, and the presence of neurofibrillary tangles. I have. In today's aging society, Alzheimer's disease is one of the social problems. No cure has been found for Alzheimer's disease, but drugs such as neurotransmitters and related substances, brain activators and cerebral circulation improvers are being developed to halt and improve the progress of the disease.
  • PET positron emission tomography
  • Examples of various biological markers include a protein called A68 having a molecular weight of 68 kD and extracted from the brain of a patient with Alzheimer's disease (Lee VL, eta1: Science, 251, 67). 5 to 678, 1991) and insoluble / ⁇ 4 protein extracted from amyloid derived from the brain of Alzheimer's disease patients (Masters CL, eta1: Proc. Natl. Aca USA, 82, 42 45-42 49, 1985), and antibodies that react with these (Wo 1 ozin BL, eta 1: Science, 23 2, 648-650, 1986) and ( ⁇ 11 sop D, etal: Neurosci Lett, 68, 252-255, 19686) have also been prepared.
  • the inventors of the present invention have studied the proteins present in the blood of Alzheimer's disease patients and the blood of normal subjects, and found that they were detected in the blood of normal subjects but not in the blood of Alzheimer's disease patients. Or, several proteins were found to be reduced in amount compared to normal individuals.
  • an antibody that recognizes the protein or a fragment of the protein is prepared, and the antibody is reacted with the blood or peripheral tissues of the subject, and the degree or presence or absence of this reaction is determined. It has been found that early detection of Alzheimer's disease can be easily performed by detecting. Furthermore, by analyzing the blood and the like of the subject by two-dimensional electrophoresis, etc., it was found that diagnosis of Alzheimer's disease can be made reliably by examining the presence or absence and the amount of these proteins.
  • the present invention was completed by producing a kit used for the former diagnosis. This invention is divided into the following four inventions.
  • [II] An antibody that recognizes the protein of [I] or a fragment of the protein.
  • [III-1] characterized by reacting a sample collected from a subject with at least one kind of antibody that recognizes the protein of [I] or a fragment of the protein, and detects the degree or presence or absence of the reaction. Diagnosis of Alheimer's disease Method.
  • [I I 1-2] A method for diagnosing Alheimer's disease, comprising analyzing a sample collected from a subject to determine the presence or amount of the protein of [I].
  • [IV] As a component, includes at least one antibody that recognizes the protein of [I] or a fragment of the protein, and a means for detecting a reaction between the antibody and a sample collected from a subject.
  • a diagnostic kit for Alheimer's disease which is characterized in that:
  • the protein of the present invention can be obtained from at least the blood of normal subjects, and is not detected in Alzheimer's disease patients or has a reduced amount as compared to normal subjects.
  • a normal person refers not only to a person who is not a patient with Alzheimer's disease, but also includes a patient with Alzheimer's disease who is in a normal state before the onset of the disease.
  • the protein of the present invention can be obtained, for example, as follows.
  • a buffer solution suitable for electrophoresis is added and sonicated, as shown in Examples described later, to be uniformly dispersed. Then, it is centrifuged under cooling to obtain a supernatant, which is used for incubation and used as an electrophoresis sample.
  • blood includes whole blood, serum, plasma, and the like.
  • Separation of this protein from the supernatant can be performed by two-dimensional electrophoresis in a conventional manner.
  • This protein of interest is determined as a spot of a protein that is clearly detected only from normal individuals by comparing it with an electrophoresis pattern obtained from blood of a patient with Alzheimer's disease by a similar method.
  • This protein was used in the examples described below. The characteristics are determined by the method shown. Subsequently, the target protein can be extracted from the two-dimensional electrophoresis gel by direct staining with coomassie, brilliant, and blue.
  • Preferred correct is a protein of this invention, for example Do not, - Anchito trypsin ( ⁇ ⁇ - protease inhibitor development b), fire! Anti-trypsin precursor, G-anti-tribsine-related protein, ⁇ , -anti-tribsine-related protein precursor, fibrinogen (eg, fibrinogen r, chain, etc.), fibrinogen precursor (eg, fibrinogen) Gen r-chain precursor, fibrinogen-chain precursor, etc.), and fragments of these proteins.
  • Antitrypsin precursor J. Mo 1. Biol., 2 1 8, 5 9 5-6 0 6 (1 9 9 9
  • ⁇ -antitrypsin-related protein precursor Ge ⁇ omics, _2_, 165-173 (1988)
  • fibrinogen r-chain precursor Biochemistry, 24, 2 0 7 7-2 0 8 6 (1 985)
  • fibrinogen chain Biochemistry, 24, 20 7 7
  • More preferred proteins of the present invention include, for example, those having the following characteristics as fragments of the above-mentioned proteins (hereinafter, these four proteins are classified into protein A in descending order of molecular weight and in descending order of isoelectric point). , protein
  • the protein of the present invention is not limited to those obtained from blood of normal persons, but may be obtained from blood other than living body, for example, from brain, cerebrospinal fluid, skin, hair, nasal mucosa, or by genetic recombination technology. Including.
  • the antibody used in the method for diagnosing Alzheimer's disease according to the present invention may be a protein or a protein which is detected in a normal person of the above [I] but not detected in Alzheimer's disease patients or whose amount is reduced as compared to a normal person. And recognizes each of the proteins described in [I] above.
  • the fragment of the protein includes a part of the amino acid sequence contained in the protein, a sugar chain modifying the protein, and the like.
  • antibodies can be either polyclonal or monoclonal and can be prepared by methods well known in the art, but monoclonal antibodies are preferred.
  • the polyclonal antibody against the protein of the present invention is suitable for animals such as egrets.
  • the resulting antiserum can be isolated and purified by subjecting the resulting antiserum to conventional procedures such as salting out ammonium sulfate, centrifugation, dialysis, and column chromatography. be able to.
  • Monoclonal antibodies can be applied to animals such as mice, for example, according to the method of Kohler & Milstein (Kohler & Milstein: Nature, 256, 495-497, 1975). After immunization by administering the protein, spleen cells are obtained from the spleen, and the cells are fused with mouse myeloma cells and the like to obtain a hybridoma. Next, the hybrid priomas producing a monoclonal antibody specific to the protein of the present invention are screened from among them, and the antibody most suitable for discriminating Alzheimer's disease patients from normal individuals from the hybrid priomas, that is, A hybridoma producing an antibody that specifically reacts with the protein of the present invention is subcloned. Then, it can be obtained by isolating and purifying from the subcloned culture supernatant or the like in the same manner as in the case of the polyclonal antibody.
  • the method for diagnosing Alzheimer's disease comprises at least one kind of a sample collected from a subject, for example, blood, skin, hair, nasal mucosa, etc., which recognizes each of the above-mentioned proteins [I] or a fragment thereof. It is characterized by reacting antibodies and detecting the degree or presence or absence of the reaction. Using the antibody obtained in the above [II], each of the proteins described in the above [I] can be diagnosed. Things.
  • antigens that specifically react with the above-mentioned antibodies present in these samples that is, are detected in normal subjects, but are not detected in Alzheimer's disease patients or have a reduced amount compared to normal subjects.
  • a protein having a fragment of the protein or the protein of the present invention It is for diagnosing Alheimer's disease.
  • this protein or a protein having a fragment of the protein is detected from the blood by the antibody of the present invention, a normal person is detected, and when not detected, Alzheimer's disease or a sign thereof is detected. It can be diagnosed that there is.
  • this protein will gradually decrease during the process of developing Alzheimer's disease in normal individuals. Therefore, a normal person is detected when a predetermined amount or more of this protein or a protein having a fragment of the protein is detected in the blood of the subject by the antibody of the present invention, and Alhiima disease or a sign thereof is detected when the amount is less than the predetermined amount. Can also be diagnosed.
  • the initial value of the amount of the protein of the present invention or a protein having a fragment of the protein in blood is measured in advance when the subject is in a normal state.
  • one or two or more of the antibodies of the present invention of the above [II], which are obtained for each of the above proteins [I] and specifically recognize the proteins, are used.
  • the antibody of the present invention is reacted directly or after pretreatment (eg, centrifugation).
  • the sample is immobilized in, for example, 10% neutral buffered formalin and then reacted with the antibody of the present invention.
  • Enzymlink Immunosorbent Atsushi ELISA
  • Enzymimnoassy EIA
  • Fluoretm Symnoassy FIA
  • Radioymnoassy RI
  • CLIA chemiluminescence immunoassay
  • CLIA chemiluminescence enzyme immunoassay
  • latex agglutination method latex agglutination method and the like.
  • the presence or amount of the protein of [I] is analyzed by analyzing a sample of a subject, and using the respective proteins described in [I] above. It can be diagnosed.
  • Examples of the analysis method at this time include an electrophoresis method (for example, a two-dimensional electrophoresis method), various chromatography methods (for example, a high-performance liquid chromatography method), a bioassay method, and the like.
  • an electrophoresis method for example, a two-dimensional electrophoresis method
  • various chromatography methods for example, a high-performance liquid chromatography method
  • a bioassay method and the like.
  • a sample of a subject or a blood sample is processed by the same method as in [I], and then subjected to two-dimensional electrophoresis. If at least one of the D spots is missing or significantly reduced (spots are significantly smaller), then Alzheimer's disease or a sign thereof can be diagnosed.
  • the kit used for the diagnosis of Alzheimer's disease in [III-11] includes at least one antibody that recognizes the protein of [I] or a fragment of the protein as a component, and the antibody and blood or It comprises a means for detecting a reaction with a sample collected from a peripheral tissue, and is obtained for each of the proteins described in the above [I].
  • Examples of the antibody used include a polyclonal antibody and a monoclonal antibody of [II], and a monoclonal antibody is preferable.
  • Methods using one type of antibody include, for example, immunohistochemical methods (eg, Examples of the method using two types of antibodies include ELISA using a sandwich method. ⁇
  • the means for detecting the reaction are those used in the detection method described in [III].
  • an enzyme-labeled anti-mouse IgG, a substrate examples include buffer solutions, blocking antibodies (eg, goat serum), and in the case of ELISA, enzyme-labeled anti-mouse IgG, substrates, carriers (eg, beads, plates, etc.), buffers, reaction stop solutions, etc. No.
  • Example 1 Pretreatment of the protein of the present invention and preparation of a sample for electrophoresis Blood was collected from Alzheimer's disease patients (5 cases, 54 to 67 years old) and normal persons (6 cases, 48 to 65 years old). , 2 volumes of sample buffer [9.9 M urine, 4% (v / v) Noridet P—40, 2.2% (v / v) Am pho 1 yte (Millipore Japan: PH 3 to: 10), 100 mM Dithiothreitol (DTT)], and sonicated three times for 5 seconds. The obtained sample was centrifuged at 20 ° C., 20 ° C. and 60 ° C. for 60 minutes, and the supernatant was incubated at 37 ° C. for 1 hour to prepare a sample for electrophoresis. . Finally, the protein concentration was about 2 mgZm1.
  • the primary electrophoresis is performed by isoelectric focusing using the Investigator 2—DE electrophoresis system (manufactured by Nippon Millipore) as the electrophoresis apparatus, and the secondary is sodium dodecyl sulfate sodium polyacrylamide electrophoresis. (SDS-PAGE) was performed.
  • the primary electrophoresis gel was prepared as follows. First, 9.5 M urea, 2.0% (V / V) Nonidet P—40, 4.1% (vZv) acrylylamidobis solution (30.8% T, 2.6% C) was added. 5. 65 ml was dispensed as a stock and stored at minus 20 ° C.
  • the primary electrophoresis was performed using 85% phosphoric acid as the anolyte and 10 N sodium hydroxide as the catholyte.
  • Overlay buffer [0.5 M urea, 0.2% (V / V) N 0 nidet P— 40, 0.1% (V / V) Am pholite, 5. OmM D TT, 0.7 M 2—Mercaptoethanol] was deposited on the plate, and the maximum voltage was set to 1500 V and the maximum current was set to 110 AZge1, and pre-electrophoresis was performed for 2 hours at room temperature.
  • the protein sample from the blood prepared in Example 1 was layered between the overlay buffer and the gel in a layer of 101, and the maximum voltage was set at 200 V and the maximum current was set at 110 AZge1. Electrophoresis was performed at room temperature until t—Hours reached 180.000 V. As an isoelectric point marker, an IEF standard (PI 4.6 to 9.6) manufactured by Biorad was used.
  • the one-dimensional gel is taken out in 10% glycerin, and the one-dimensional gel equilibration buffer (0.3 MT ris Base, 0.075 MT ris HC 1,3) is taken for 2 minutes. After equilibration in 5% sodium dodecyl sulfate (SDS), 5 OmM DTT, 0.1% bromphenol blue), they were placed on a secondary electrophoresis gel.
  • SDS sodium dodecyl sulfate
  • 5 OmM DTT 0.1% bromphenol blue
  • Second electrophoresis gel is D uracr y 1 (manufactured by Nippon Miripore) 2 96 lml, Tris buffer (130.8 g / 1 Tris Base, 66.3 g / l Tris HC1) 2 20.5 ml, M i 1 1 i— Q water 3 6 5.4 ml, 10 3 ⁇ 4 SDS 9. O ml, tetramethylethylenediamine (TEMED) 0.447 m 1, 10% APS 2. 24 ml was mixed and made to a thickness of 1 mm between 26 x 26 cm glass plates.
  • the two-dimensional electrophoresis buffer was adjusted with the composition of 25 mM Tris Base, 19 2 mM glycine, 0.1% SDS, and the electrophoresis was performed at 20 ° C at a maximum voltage of 500 V and maximum power. The procedure was performed at 160,000 mW / ge 1 until the stained surface was 1 cm from the bottom of the gel.
  • SDS-PAGE standard Low, MW 1440000-974 manufactured by Biorad was used.
  • Example 2 (2) The secondary electrophoresis gel obtained in Example 2 (2) was stained with silver by a conventional method.
  • the blotting membrane was stained with PhastGelBluR (Pharmacia) to cut out the target spots.
  • PhastGelBluR PhastGelBluR
  • amino acid sequence of these proteins from the ⁇ -terminal to the 12th to 15th residues is, as shown in the sequence listing below,
  • Protein G 1 u-Asp p o -Gln-Gly -Asp -Ala-15
  • H is Protein BA sp— A la— X aa— G in — X aa— A sp 1
  • Xaa is an unidentified amino acid, but
  • X a a of the third residue of B is A 1 a
  • X a ay5 of the sixth residue is T h r
  • Example 2 (2) Separation of water-soluble proteins in blood using high-resolution two-dimensional electrophoresis was performed in the same manner as in Example 2 (1), followed by primary electrophoresis, and then in Example 2 (2). Then, secondary electrophoresis was performed to obtain a two-dimensional electrophoresis gel.
  • the blotting membrane was incubated for 24 hours at room temperature with a 100-fold diluted antibody (primary antibody) described below. After washing three times with TBBS, the cells were incubated for 1 hour and 30 minutes with alkaline phosphatase-labeled goat anti-Peacock IgG antibody (secondary antibody). After washing three times with TBBS, an alkaline phosphatase reaction was performed on the basis of W estern Blue AP-substrate solution (Promega, fr S3841) until protein spots appeared. Washed three times with TBBS.
  • the image of the colored block film is captured by a scanner, and each plot is converted to an optical density, so the optical intensity per dot 1/300 inch square is a relative value from 0 to 255. By multiplying this by the area of each block, the color intensity of each block as a whole was quantified.
  • antibody (I) (Hereinafter referred to as antibody (I))
  • antibody (II) (Hereinafter referred to as antibody (II))
  • the spot of the target protein was specifically and strongly stained in a sample of a normal subject.
  • some spots with slightly higher molecular weight were detected, which are considered to be precursors of the target protein or similar.
  • Spots of this protein were stained very faintly in samples from patients with Alzheimer's disease, but no significant differences were found in the other spots. Therefore, the target protein is considered to be ⁇ ] -antitrypsin itself or a fragment thereof.
  • the spot of the target protein was specifically and strongly stained in a sample of a normal subject.
  • some spots with slightly higher molecular weight and more alkali-bias were detected, which are considered to be precursors of the target protein or similar.
  • spots of this protein of interest were stained very faintly or almost unstained, while the other spots showed large individual differences and no significant differences were found. Therefore the target protein The quality of the protein is considered to be fibrinogen itself or a fragment thereof.
  • Blood collected from patients with normal Alzheimer's disease and normal subjects is diluted 2-fold with the same sample buffer as in Example 1, and incubated at 37 ° C for 1 hour. Samples were washed with TBS (20 mM Tris, 500 mM sodium chloride, pH 7.4, 0.05% sodium azide)-5% sodium dodecyl sulfate (SDS). Dilute 5-fold, mix gently, and incubate at 37 ° C for 2 hours. After incubation, centrifuge the sample at more than 700,000 xg to remove insoluble proteins as pellets. After setting the nitrocellulose membrane immersed in TBS in the slot block device to make it a suction state, immediately perform the operation of dispensing the sample (200 ⁇ 1) to each cell twice.
  • TBS 20 mM Tris, 500 mM sodium chloride, pH 7.4, 0.05% sodium azide
  • SDS sodium dodecyl sulfate
  • washing buffer 3 times diluted with 0.1% Tween 20 containing TBS
  • Transfer ⁇ to the appropriate primary antibody dilute in wash buffer, and incubate overnight at room temperature.
  • the antibody is used after diluting 500-fold with a washing buffer.
  • the membrane is washed three times for 10 minutes.
  • Immerse the membrane in a secondary antibody algal phosphatase-labeled goat anti-mouse IgG antibody
  • a secondary antibody algal phosphatase-labeled goat anti-mouse IgG antibody
  • the membrane is washed three times for 10 minutes.
  • the membrane is further rinsed easily with TBS and the substrate solution (5-bromo-4-chloro-3-indolinolephosphate (15 mg), nitrobenzotetrazolium chloride (30 mg), 0.1 M sodium bicarbonate, 1.O mM Transfer to magnesium chloride, pH 9.8) to develop color.
  • the colored membrane is then washed with steam for several minutes.
  • the coloring intensity of each blot was quantified.
  • antibody ( ⁇ ) (Hereinafter referred to as antibody ( ⁇ ))
  • a diagnostic kit having the above configuration is manufactured.
  • the protein of the present invention can be obtained from at least the blood of normal subjects and is detected in normal subjects, but is not detected in Alzheimer's disease patients or its amount is reduced as compared with normal subjects. Therefore, antibodies useful for diagnosing Alzheimer's disease can be prepared using this protein. Furthermore, this protein is expected to be very useful for research such as investigation of the cause of Alzheimer's disease.
  • the antibody of the present invention recognizes the protein of the present invention or a fragment thereof present in the blood or peripheral tissues of a subject and reacts with the protein or a protein having a fragment of the protein. By examining it, it is useful for diagnosing Alzheimer's disease.
  • a sample collected from blood, peripheral tissues, or the like can be used, so that diagnosis of Alzheimer's disease can be easily performed. Furthermore, the antibody is reacted with the sample, and not only the presence or absence but also the degree of the reaction is detected, so that early diagnosis of Alzheimer's disease can be made.
  • a sample collected from blood, peripheral tissues, or the like is analyzed to determine the presence or amount of the protein of the present invention. Not only can it be done Pre-diagnosis can also be performed. According to the kit for diagnosing Alzheimer's disease of the present invention, Alzheimer's disease can be easily diagnosed. Sequence Listing SEQ ID NO: 1 Sequence length: 15 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Fragment type: N-terminal fragment sequence

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Abstract

Substance permettant de diagnostiquer la maladie d'Alzheimer et d'étudier la cause et le traitement de celle-ci, telles qu'une protéine présentant les caractéristiques exposées ci-après et un anticorps produit par la protéine ou des fragments de celle-ci, procédé de diagnostic de la maladie d'Alzheimer et kit prévu à cette fin: (1) poids moléculaire: 40 kD, déterminé par électrophorèse sur gel de polyacrylamide au dodécylsulfate de sodium; (2) point isoélectrique: pI = 8,2; (3) séquence d'acides aminés (A) commençant par la terminaison N jusqu'au 15e résidu, où Xaa représente un résidu d'acide aminé non identifié.
PCT/JP1994/001359 1993-08-17 1994-08-17 Procede de diagnostic de la maladie d'alzheimer WO1995005395A1 (fr)

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AU74670/94A AU7467094A (en) 1993-08-17 1994-08-17 Method of diagnosing alzheimer's disease

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JP5/203449 1993-08-17
JP20344993 1993-08-17

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6045773A (en) * 1995-10-17 2000-04-04 G.D. Searle & Co. Method of detecting cyclooxygenase-2

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4839283A (en) * 1986-12-30 1989-06-13 Zymogenetics, Inc. Method of expressing alpha-1-antitrypsin in yeast
JPH0418100A (ja) * 1990-05-09 1992-01-22 Mitsui Toatsu Chem Inc 神経栄養活性抑性物質
WO1992001464A1 (fr) * 1990-07-19 1992-02-06 The Scripps Research Institute Inhibition de la liaison entre le recepteur mac-1 et le fibrinogene a l'aide des homologues de d30

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4839283A (en) * 1986-12-30 1989-06-13 Zymogenetics, Inc. Method of expressing alpha-1-antitrypsin in yeast
JPH0418100A (ja) * 1990-05-09 1992-01-22 Mitsui Toatsu Chem Inc 神経栄養活性抑性物質
WO1992001464A1 (fr) * 1990-07-19 1992-02-06 The Scripps Research Institute Inhibition de la liaison entre le recepteur mac-1 et le fibrinogene a l'aide des homologues de d30

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ADV. EXP. MED. BIOL., Vol. 281, 1990, DOMINIC W. CHUNG, "Nucleotide Sequences of the Three Genes Coding for Human Fibrinogen", pages 39-48. *
BIOCHEMISTRY, Vol. 24, No. 8, 1985, MARK W. RIXON, "Nucleotide Sequence of the Gene for the gamma Chain of Human Fibrinogen", pages 2077-2086. *
GENOMICS, Vol. 2, No. 2, 1988, JIA-JU BAO, "Molecular Structure and Sequence Homology of a Gene Related to alpha1-Antitrypsin in the Human Genome", pages 165-173. *
J. MOL. BIOL., Vol. 218, No. 3, 1991, U. BAUMANN, "Crystal Structure of Cleaved Human alpha1-Antichymotrypsin at 2.7A Resolution and Its Comparison With Other Serpins", pages 595-606. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6045773A (en) * 1995-10-17 2000-04-04 G.D. Searle & Co. Method of detecting cyclooxygenase-2

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