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WO1997008549A1 - Procede de detection d'affections renales, produit et kit de diagnostic appropries - Google Patents

Procede de detection d'affections renales, produit et kit de diagnostic appropries Download PDF

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Publication number
WO1997008549A1
WO1997008549A1 PCT/JP1996/002461 JP9602461W WO9708549A1 WO 1997008549 A1 WO1997008549 A1 WO 1997008549A1 JP 9602461 W JP9602461 W JP 9602461W WO 9708549 A1 WO9708549 A1 WO 9708549A1
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WIPO (PCT)
Prior art keywords
integrin
antibody
renal disease
urine
complex
Prior art date
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PCT/JP1996/002461
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English (en)
Japanese (ja)
Inventor
Naomasa Yamamoto
Kenjiro Tanoue
Koichi Utsumi
Hiroshi Saito
Yasuhiro Katagiri
Masaharu Kotani
Shuichi Miyaura
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Tokyo Metropolitan Institute of Medical Science
Seikagaku Corp
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Tokyo Metropolitan Institute of Medical Science
Seikagaku Corp
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Application filed by Tokyo Metropolitan Institute of Medical Science, Seikagaku Corp filed Critical Tokyo Metropolitan Institute of Medical Science
Priority to JP51013097A priority Critical patent/JP3870242B2/ja
Publication of WO1997008549A1 publication Critical patent/WO1997008549A1/fr
Anticipated expiration legal-status Critical
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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

Definitions

  • the present invention relates to a method for detecting a renal disease, a diagnostic agent and a diagnostic kit, and more particularly, to a method for measuring a renal disease by measuring integrin ⁇ 3 in a body fluid. It relates to the method of detecting BACKGROUND ART
  • diagnosis has been made based on examination of leakage of blood components (cells such as red blood cells and white blood cells and plasma proteins) into urine by urine sediment, urine protein and the like.
  • urine / 3 2 - concentration microglobulin (microglobulin) and N- ⁇ cetyl glucosaminoglycans NIDA one peptidase (NAG) is to show the extent of injury of tubular.
  • the activated complement system is thought to deposit on glomerular epithelial cells and damage the glomerulus, resulting in urinary complement. Detection of a component or measurement of its activity has been referred to.
  • the activated complement system binds to vitronectin (VN), a plasma protein that inactivates it, and forms a VNZCb5-9 complex to be excreted in urine. Measuring vitronectin has also been one method of diagnosing renal disease.
  • Enzymim Assay A method is known in which a sample urine is added to a plate for EIA), and then the antibody is added to measure normal ⁇ tissue antigen in the urine to diagnose renal injury (JP-A-63-112994).
  • integrin is an adhesion molecule on the surface of a cell membrane in which a complex of one subunit and three subunits forms a one-to-one complex
  • various types of molecules are known from combinations of different subunits and three subunits.
  • ⁇ subunit against the / 3 3 Sabuyunitto £ ⁇ and 0: 1 of 2 types are known Till.
  • I i b 3 3 Complex known to be a major integrin present on platelets, alpha [nu 3 3 double coalescence is known to be a receptor for vitronectin, also a the tissue by connexion yarn ball body such as immunostaining It has been reported that the vy S 3 complex is frequently found (N mark hron, 68 (1994) p. 87-96). It is known that the urine of patients with renal disease excretes platelets and cells from the kidney, such as glomerular cells, but integrin 3 is present in the body fluids of patients with renal disease, specifically, urine. Its presence and the association between bodily fluids, specifically urinary integrins, and renal disease have not been studied.
  • the present invention has been made in view of the above, and is a method for easily detecting the presence or absence of renal disease, particularly glomerular injury, or the degree of renal tissue damage. It is an object to provide a diagnostic agent and a diagnostic kit.
  • the present inventors have found extensive research result, the body fluid of a patient having renal disease, the presence of integrin / 8 3 More specifically urine, and more specifically in the urine sediment in order to solve the above problems and, finding a way to measure this integrin 3 3 easily and accurately, dull to the present invention.
  • detection method preferably urine, more preferably renal disease and measuring the integrin yS 3 in urine sediment,
  • a solid phase body to which an antibody against 3 integrin) 3 3 is fixed a kit for diagnosing renal disease comprising the antibody labeled with or capable of being labeled with a labeling substance
  • antibodies against I integrin yS 3 (integrin ⁇ 3 and antibodies specifically reactive, hereinafter referred to as "anti-integrin / 3 3 antibody” ) By an antigen-antibody reaction.
  • a specific method for measuring the integrin / 3 3 by antigen-antibody reaction for example, subjected to electrophoresis Solubilized urine sediment as a specimen, followed by transferring the analyte after electrophoresis to a membrane, anti-integrin on the film; S 3 antibody was added, integrase phosphorus by antigen-antibody reaction; and measuring the S 3, the integrin urinary sediment was solubilized, anti integrin Glynn; is reacted with S 3 antibody, antibody has one integrin / 3 3 - antibody and a method of measuring the integrin y3 3 by detect the formation of the complex, but is not limited thereto.
  • a kidney disease refers to a disease associated with damage to kidney glomeruli and the like, such as nephritis and nephropathy (nephrosis).
  • Such renal diseases are known to be caused by various causes, such as those caused by binding or deposition of immune complexes. Specifically, for example, systemic lupus erythematosus (SLE), IgA ⁇ disease , Diabetic nephropathy, membranous nephropathy, hypertensive renal sclerosis, membranous proliferative nephritis, acute glomerulonephritis, focal glomerulosclerosis, purpura nephritis and the like.
  • SLE systemic lupus erythematosus
  • IgA ⁇ disease Diabetic nephropathy, membranous nephropathy, hypertensive renal sclerosis, membranous proliferative nephritis, acute glomerul
  • Integrin S 3 of the present invention of the integrin; molecule having a 3 3 Sabuyunitto (alpha [nu S 3 complex, nb / 8 3 complex, etc.),; as 3 sub Buyunitto 3 3 Sabuyunitto itself, And fragments thereof.
  • Integrin / 93 is excreted in the urine of patients with renal disease, but is not present in the urine in a soluble form such as urine protein, but is excreted in the urine. It is thought that it exists in a form bound to fragments. That is, the integrin / 3 3 to be discharged into diuresis by the injury of glomeruli like kidney, measured as injury marker ⁇ tissue Thereby, renal disease can be detected. This detection of renal disease includes the determination of the presence or absence and degree of renal disease, as well as prognostic determination such as evaluation of therapeutic effects.
  • Anti-integrin used in the present invention the 9 3 antibody, / 3 3 subunits, three sub Buyuni' integrin molecule comprising Bok ( ⁇ complex Ya ⁇ composite) or antibodies such as recognizing and these fragments Can be mentioned.
  • alpha [nu / 3 3 complex (glomerular origin) of ⁇ 3 Sabuyunitto and a I Ib; S 3 complex (derived platelets) of; 6 3 follows the line if you wish to measure and differentiate between Sabuyunitto Just do it. That a, lb; 8 3 a l lb Sabuyuni' Bok specifically reactive with monochromatic one monoclonal antibody use the t, was a by antigen-antibody reaction complex (platelet-derived),! »Sabuyunitto amount measured to determine the 3 3 Sabuyunitto amount of platelet-derived. Then by subtracting the / 3 3 Sabuyunitto amount from platelet from all / 3 3 Sabuyunitto amount can be determined integrin> 3 3 content derived from glomeruli.
  • the method for detecting a renal disease of the present invention uses a body fluid, preferably urine, more preferably a urine sediment or a solubilized urine sediment (hereinafter, unless otherwise specified, collectively referred to as “urine sediment”) as a specimen, It is performed by measuring 9 3; integrin in the sample.
  • a body fluid preferably urine, more preferably a urine sediment or a solubilized urine sediment (hereinafter, unless otherwise specified, collectively referred to as “urine sediment”) as a specimen, It is performed by measuring 9 3; integrin in the sample.
  • a method for measuring integrin ⁇ 3 in urine sediment there is a method in which an anti-integrin;
  • Anti-integrin used in the present invention the S 3 antibody, modified / 3 3 Sabuyunitto, a v yS 3 antibody has high reactivity with complex or a iib S 3 complexes are preferred Te cowpea in unmodified or solubilized .
  • anti-integrin 83 antibodies for example, Ant i -Human Integrin ⁇ (clone number VNR3, VNR5, Takara Shuzo Co., Ltd.) can be used such as , prepared monoclonal antibody of the present inventors (2 J 40 antibody, see example), even in the native / 3 3 sub Yunitto, also modified with sodium dodecyl sulfate (SDS) / 3 3 Sabuyuni Tsu DOO Is particularly preferred because of its high reactivity.
  • SDS sodium dodecyl sulfate
  • anti-integrin 5 3 antibodies can be also child prepared as below, for example according to a conventional method.
  • Purified 3 3 Sabuyunitto used as an antigen it is possible to obtain in a known manner.
  • normal human platelets are solubilized and converted to a peptide having the amino acid sequence of Arg-Gly-Asp or a peptide having the amino acid sequence of Gly-Arg-Gly-Asp-Ser_Pro-Lys (SEQ ID NO: 1).
  • Poly Skr Norre anti-integrin S 3 antibodies such as mice, rats, guinea pigs, Usagi, catcher formic
  • the animals to be immunized such as Hijji immunized with the above antigens may be obtained by removing a serum from these animals . It is preferable to use an auxiliary agent (adjuvant) in immunizing the animal to be immunized, since it activates antibody-producing cells. From the obtained antiserum, the immunoglobulin fraction may be purified by a conventional method.
  • a monoclonal anti-integrin antibody can be obtained, for example, as follows. That is, the antigen is administered intraperitoneally, subcutaneously, or to the footpad of an animal to be immunized, such as a mouse, rat, guinea pig, rabbit, goat, or sheep, and then the spleen or popliteal lymph node is removed. The cells collected from these cells are fused with myeloma cells, which are tumor cell lines, to establish hybridomas. The resulting hybridomas are continuously grown, and the specific antibodies against the above antigens are continuously obtained from the obtained hybridomas. Select the cell line that will produce. By culturing the selected strain in a suitable medium, a monoclonal anti-integrin antibody can be obtained in the medium. Alternatively, by culturing the High Priestess dormer in vivo, such as mouse peritoneal cavity it can be mass production of monoclonal anti Integurin [delta] 3 antibody.
  • Cells used for cell fusion include lymph node cells and peripheral blood Lymphocytes and the like can be used. Further, the myeloma cell line is preferably derived from the same cell line as compared to that derived from a heterologous cell type, and a stable antibody-producing hybridoma can be obtained.
  • Methods for purifying the obtained polyclonal and monoclonal antibodies include salting out with sodium sulfate, ammonium sulfate, etc., low-temperature alcohol precipitation, selective precipitation fractionation using polyethylene glycol or isoelectric point, electrophoresis, DEAE Ion-exchange chromatography using an ion exchanger such as tylaminoethyl) mono-derivative and CM (carboxymethyl) mono-derivative; affinity chromatography using protein A or protein G; Examples include tight chromatography, immunosorbent chromatography in which an antigen is immobilized, gel filtration, ultracentrifugation, and the like.
  • anti-integrin; S 3 antibodies can be used as it is, it is also possible to use those obtained by Furagume cement of. Preservation of the antibody's antigen binding site (F ab) during antibody fragmentation is essential for binding the antigen to the antibody, so proteases that do not degrade the antigen binding site (eg, plasmin, pepsin, papain) Etc.), a fragment containing Fab obtained by treating the antibody can also be used.
  • fragment preparative Ya chimeric antibodies e.g., anti Integurin including genetic engineering F ab; 8 3 antibody F chimeric antibodies containing an ab portion
  • fragments and chimeric antibodies can also be used in the present invention.
  • anti-integrin 3 3 antibody forms of the antigen-antibody reaction using the anti Integurin / 9 3 antibody as above is not particularly limited, the measurement system used for ordinary Imunoassi (immunoassays) can be applied to the present invention regardless of the type.
  • the measurement system include an immunoblotting method (for example, a Western plotting method), a labeled immunoassay method (for example, an EIA method, an enzyme-linked immunosorbent assay (ELISA method), and a radioimmunoassay). Assay method, fluorescence immunoassay method, etc.), flow cytometry method and the like. It is.
  • These measurement methods are known methods [S. Irie, edited by "Radio Simnoassy",
  • electrophoresis is performed using a body fluid, preferably urine, more preferably a solubilized urine sediment as a sample, and then the sample after electrophoresis is transferred to a membrane.
  • the anti-integrin 3 antibody is added, can be done by measuring the integrin) S 3 by antigen-antibody reaction.
  • the electrophoresis is preferably performed by SDS-polyacrylamide gel electrophoresis.
  • the conditions for SDS-polyacrylamide electrophoresis and the conditions for transfer to a membrane can be appropriately determined by those skilled in the art according to the known conditions used for the Western blotting method.
  • the membrane may be a polyvinylidene difluoride (PVDF) membrane, a nitrocellulose membrane, a nylon membrane, or the like.
  • PVDF polyvinylidene difluoride
  • Measurement of the antigen-antibody reaction in the Western blotting technique leave labeled in advance with a labeling substance anti integrase phosphorus 3 3 antibodies, and Inkyubeto the labeled antibody and the membrane, the labeled antibody bound to the integrin / [delta] 3 of the H ⁇
  • the detection can be performed by detecting the labeling substance by a method suitable for the labeling substance.
  • anti-integrin unlabeled; and S 3 Inkyubeto antibody the membrane and, Integurin; 5 3 anti Integurin; after forming the conjugates of S 3 antibody, anti-integrin; be used in the preparation of 3 3 antibody
  • a labeled secondary antibody obtained by labeling an antibody against immunoglobulin of an immunized animal (secondary antibody) with a labeling substance is bound to the conjugate, and the labeling substance in the secondary antibody bound to the conjugate is detected.
  • the antigen-antibody reaction can be measured. Is the anti-integrin 5 3 antibody was modified with the modifying agent (eg SDS, etc.); 3 3 Sabuyuni' antibody is highly reactive to Bok is preferred.
  • Labeling immunoassay for example Sanditsuchi EI Alpha method, a body fluid, preferably urine, is Ri preferably good integrin urine sediment was solubilized; and 9 3 is reacted with anti-integrin 3 3 antibodies, sandwich complex , i.e. anti-integrin; 8 3 antibody has one Integrated Darrin; 9 3 - can and this carried out by detecting the formation of a complex of anti-integrin / 3 3 antibody.
  • the complex is formed by using a labeled antibody as one of the antibodies forming the complex and using an antibody capable of binding or binding to the solid phase on the other side, and using the labeled substance in the complex as the labeled substance. Can be detected by the method described above.
  • the labeled secondary antibody is bound to the complex, and the labeling substance of the labeled secondary antibody bound to the complex is measured to form the complex. Can be detected. More specifically, examples of the sandwich EIA method include the following methods, but are not limited thereto.
  • the solid phase body examples include a microtiter plate (imnoplate), a ball of polystyrene, latex particles, a test tube wall, and a metal colloid.
  • a method for immobilizing an antibody on these solid phase bodies include a physical adsorption method and a covalent binding method. 75, published by Kodansha, pp. 9-75).
  • the physical adsorption method is preferred because it is simple and widely used as a method for fixing an antibody to a solid phase.
  • the portion to which the antibody is not bound may be blocked by serum albumin, gelatin, milk protein, or the like.
  • the solid phase and the liquid phase are separated, and the integrin in the specimen sample can also be measured by measuring the labeled substance in the liquid phase. it can.
  • the integrin / S 3 labeled with a labeling substance, anti-integrin was immobilized on a solid phase with a test sample; mixed with 9 3 antibody to measure the amount of labeled substance bound to the immobilized antibody, so-called competitive method as well, integrin specimen sample; it is possible to measure the 3 3.
  • the antigen-antibody reaction is not limited to the solid phase method as described above, and may be performed by a so-called liquid phase method without using an immobilized antibody. That is, anti-integrin; mixing 3 3 antibody and the test sample, integrin Glynn yS 3 was further added a labeled secondary antibody - anti-integrin; to precipitate the complex of 9 3 antibody has one labeled secondary antibody, the precipitation This is a method for detecting the contained label.
  • the labeling substance used for labeling the antibody enzyme (Peruokishidaze, alkaline phosphatase, ⁇ , such as single-galactosidase), a radioactive isotope (1 25 1, 1 3 1 I, 3H , etc.), fluorescent substances (Full O receptacle in Isothiocarbazate Xia And chemiluminescent substances (such as noreminol), and other substances (such as biotin and avidin).
  • enzyme Peruokishidaze, alkaline phosphatase, ⁇ , such as single-galactosidase
  • a radioactive isotope (1 25 1, 1 3 1 I, 3H , etc.
  • fluorescent substances Flul O receptacle in Isothiocarbazate Xia And chemiluminescent substances (such as noreminol)
  • other substances such as biotin and avidin.
  • the method of labeling the antibody may be a known method suitable for the labeling substance, for example, when labeling an enzyme.
  • the glutaraldehyde method the periodic acid crosslinking method, the maleimide crosslinking method, the carpoimide method, and the like.
  • the chloramine T method the lactoperoxidase method, etc. (Seismic Chemistry Laboratory Course 5 Immunobiochemistry Research method, Tokyo Kagaku Dojin, published in 1986).
  • the urinary sediment is not particularly limited as long as it is insoluble in urine.
  • insoluble proteins, cells, cell fragments, etc. in urine are separated by centrifugation or ultrafiltration. What was concentrated by a membrane etc. is mentioned.
  • centrifugal separation is simple and therefore preferable.
  • the centrifugation conditions include, for example, the conditions of centrifugation at 400 X g or more for 5 minutes or more.
  • a urinary sediment is solubilized by adding a surfactant, a denaturant (solubiliser) such as guanidine hydrochloride or urea that denatures proteins.
  • the denaturant (solubiliser) is preferably a surfactant, and more preferably an ionic surfactant.
  • an alkyl sulfate such as sodium dodecyl sulfate (SDS) is used among ionic surfactants, its concentration is, for example, about 0.1 to 5% based on urine sediment.
  • a urine sediment solubilized by adding a surfactant or the like is usually used as a specimen sample.
  • the type and concentration of the surfactant can be appropriately selected within a range that does not affect the antigen-antibody reaction.
  • surfactants include polyoxetylene alkylphenyl ethers (Triton-based surfactants) (Triton X-100, etc.), polyoxyethylene sorbitan alkyl esters (Tween-based surfactants) c body fluid non-ionic surfactant such as agent) and the like, preferably urine, more preferably in the urinary sediment integrin / 3 3 of quantification, using a known amount integrin) 3 3 as a standard substance
  • the calibration can be performed by preparing a calibration curve based on the relationship between the concentration of the standard substance and the amount of the detected label, and comparing with the calibration curve. In the competition method, a certain amount of standard substance is added to the antigen-antibody reaction system.
  • integrin s 3 With unlabeled integrin s 3 of a known amount is added, the relationship between the amount of label is detected to a standard concentration Nitsu L, Te it is sufficient to create a calibration curve.
  • anti-integrin by measuring the integrin in body fluids by using a 3 3 antibody, it is possible to perform detection of renal disease. That is, anti-integrin Glynn; 9 3 antibodies can be used as a diagnostic agent for renal diseases.
  • a diagnostic kit may be added by adding substances, solubilizing agents (denaturing agents) that do not affect the antigen-antibody reaction. Solid-phase body may be allowed to secure the advance anti-integrin / 9 3 antibody, and favored arbitrary. By using such a kit, integrin 3 can be easily measured.
  • the affinity column is a CNBr-activated Sepharose (manufactured by Pharmacia, Inc.) to which the Gly-Arg-Gly-Asp_Ser-Pro-Lys (SEQ ID NO: 1) peptide is used as a ligand, and this peptide is bound in a conventional manner. A column packed with)) was used.
  • This complex was identified as two nodes by silver staining after SDS-polyacrylamide gel electrophoresis. Obtained a l lb / S 3 complex in SDS- polyacrylamide gel electrophoresis, alpha ,, b Sabuyunitto and; fractionated separated fraction into a 8 3 Sabuyunitto was isolated yS 3 Sabuyunitto.
  • a solution containing promophenol blue hereinafter, abbreviated as “solubilizing solution”) 400 ⁇ 1 was added and dissolved (solubilized), and heated at 100 for 5 minutes.
  • the obtained urine sediment sample was stored at 20 ⁇ until measurement, and dissolved and used at the time of measurement.
  • SDS-polyacrylamide gel electrophoresis was performed with 20 urine sediment samples per lane.
  • the electrophoresis was performed using Resep Ge 1 (5-20% gradient) (manufactured by Washimori Mori Co., Ltd.) at 30 mA for about 1 hour until the bromophenol buffer reached the tip of the gel.
  • proteins were transferred from the gel after electrophoresis using Immobilon TM (PVDF membrane, Millipore ⁇ ⁇ ) according to a conventional method.
  • Monoclonal antibody 2J40 (1: 400 dilution) was added to the membrane after the transfer, and incubated at room temperature for 1 hour. Washing was performed twice with PBS—0.05% Tween 20 for 5 minutes and then for 10 minutes.
  • HRP horseradish peroxidase
  • the above-mentioned healthy subjects and patients with renal disease were 23 healthy subjects and 48 patients with various renal diseases.
  • the breakdown of the disease was 19 cases of systemic lupus erythematosus (SLE) (6 cases of active (acute), Active (chronic) 13 cases), Ig A nephropathy 10 cases, diabetic nephropathy (DM) 6 cases, minimal change nephropathy 6 cases, membranous nephropathy (MN) 4 cases, membranous proliferative
  • SLE systemic lupus erythematosus
  • DM diabetic nephropathy
  • MN membranous nephropathy
  • MPGN membranous nephropathy
  • HTN hypertensive renal sclerosis
  • the healthy subjects were 13 males and 10 females, aged between 22 and 59 years.
  • Patients with various renal diseases included 20 males and 28 females, ages 23-68.
  • Table 1 shows the results.
  • Table 1 Urinary integrin; 3 3 of excretion
  • integrin S3 was hardly excreted in urine in healthy subjects.
  • quantitative results of integrin ⁇ 3 showed that its S output varied depending on the type of kidney disease, and also varied depending on the activity in the same kidney disease group. That is, the amount of urinary integran ⁇ 3 was much higher in active SLE than in inactive 3 £ compared to 3 inactive. Also, slight changes in which no histopathological glomerular injury was observed in nephropathy, integrin from urinary sediment; 3 3 was detected.
  • P 2 antibody as a comparative example ( "M b / S 3 A monoclonal antibody against the complex, Cosmo Bio) was added to each, and a FITC-labeled secondary antibody was further added.
  • Flow cytometry was performed using a FACS can (BECTON DICKINSON) according to a conventional method. It was. As a result, 2 J 40 antibodies and cells, etc. of the urine sediment for LM609 antibody stained positively, although integrin / 3 3 was detected, there was negative for P2 antibodies. platelet derived alpha, , b 3 3 complexes were not detected.
  • eXAMPLE 3 anti-integrin was prepared by the method described in preparation> 3 3 antibody (2 J 40 antibodies) phosphate buffered saline (PH7.2 ⁇ 7.5 , Without divalent ion) (hereinafter referred to as PBS (—) ), Add 50 1 (lg / ⁇ ) of this solution to each well of Imnoplate (trade name: Maxisorp, manufactured by Nunc), and store at 4 for 16 hours. Coated evenly.
  • the plate is then washed twice with PBS (-), and 3% Pseudoserum albumin (BSA) (BSA) is used as a blocking substance to cover the parts of the wells that are not coated with anti-integrin / 333 antibody. (Industry Co., Ltd.) containing PBS (-) solution was added and left at room temperature for 2 hours.
  • PBS Pseudoserum albumin
  • a body fluid preferably urine, more preferably urine sediment, very preferably
  • solubilizers e.g., T Riton based or T ween surfactant
  • integrin e.g., T Riton based or T ween surfactant

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Abstract

La présente invention concerne la détection des affections rénales par dosage d'intégrine β3 dans un sédiment urinaire solubilisé au moyen d'une technique d'immunotransfert de type Western, un procédé de dosage immunologique à marquage etc., conjointement avec l'utilisation d'un anticorps dirigé contre l'intégrine β3.
PCT/JP1996/002461 1995-08-31 1996-08-30 Procede de detection d'affections renales, produit et kit de diagnostic appropries Ceased WO1997008549A1 (fr)

Priority Applications (1)

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JP51013097A JP3870242B2 (ja) 1995-08-31 1996-08-30 腎疾患の検知法、診断薬及び診断用キット

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JP22375595 1995-08-31

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002037099A1 (fr) * 2000-10-27 2002-05-10 International Reagents Corporation Procede de diagnostic de la nephropathie
JP2008122410A (ja) * 2000-10-27 2008-05-29 Masanori Hara 腎障害の検査方法
WO2009041577A1 (fr) * 2007-09-27 2009-04-02 Niigata University Agent de prétraitement d'urine pour la détermination de protéine urinaire, procédé de prétraitement d'urine et procédé de détermination de protéine urinaire
WO2014023819A1 (fr) * 2012-08-10 2014-02-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Procédés de prédiction de la durée de survie d'un patient souffrant d'un glioblastome
JP2014517276A (ja) * 2011-05-13 2014-07-17 キングス・カレッジ・ロンドン 血小板感受性に関係する方法と組成物
CN115667921A (zh) * 2020-05-26 2023-01-31 东洋纺株式会社 尿沉渣用染色液

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JPS63112994A (ja) * 1986-10-31 1988-05-18 Hajime Inamoto モノクロ−ナル抗体を用いた尿中腎抗原の測定による腎障害の診断方法

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JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, Vol. 6, No. 3 (1995), UTSUMI K.; YAMAMOTO N.; KATAGIRI Y.; KOTANI M.; KANEKO T.; UJIIE K.; NAGAI T.; IINO Y.; TERASI A.; SAITO H.; TANOUE, "Urinary excretion of Vitronectin (VN) Receptor (alpha-V-beta-3 Integrin) as a New Markers for Glometrular Injuries", p. 433. *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002037099A1 (fr) * 2000-10-27 2002-05-10 International Reagents Corporation Procede de diagnostic de la nephropathie
US6969591B2 (en) 2000-10-27 2005-11-29 Masanori Hara Method for diagnosing nephropathy
JP2008122410A (ja) * 2000-10-27 2008-05-29 Masanori Hara 腎障害の検査方法
WO2009041577A1 (fr) * 2007-09-27 2009-04-02 Niigata University Agent de prétraitement d'urine pour la détermination de protéine urinaire, procédé de prétraitement d'urine et procédé de détermination de protéine urinaire
EP2196803A4 (fr) * 2007-09-27 2010-12-22 Univ Niigata Agent de prétraitement d'urine pour la détermination de protéine urinaire, procédé de prétraitement d'urine et procédé de détermination de protéine urinaire
JPWO2009041577A1 (ja) * 2007-09-27 2011-01-27 国立大学法人 新潟大学 尿中タンパク質定量用の尿前処理剤、尿前処理方法、及び尿中タンパク質定量方法。
US8105840B2 (en) 2007-09-27 2012-01-31 Niigata University Urine pretreatment agent for urinary protein quantitation, urine pretreatment method, and urinary protein quantitation method
JP2014517276A (ja) * 2011-05-13 2014-07-17 キングス・カレッジ・ロンドン 血小板感受性に関係する方法と組成物
WO2014023819A1 (fr) * 2012-08-10 2014-02-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Procédés de prédiction de la durée de survie d'un patient souffrant d'un glioblastome
CN115667921A (zh) * 2020-05-26 2023-01-31 东洋纺株式会社 尿沉渣用染色液

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