[go: up one dir, main page]

WO2009088022A1 - Nouveau marqueur de cancer et diagnostic l'utilisant - Google Patents

Nouveau marqueur de cancer et diagnostic l'utilisant Download PDF

Info

Publication number
WO2009088022A1
WO2009088022A1 PCT/JP2009/050088 JP2009050088W WO2009088022A1 WO 2009088022 A1 WO2009088022 A1 WO 2009088022A1 JP 2009050088 W JP2009050088 W JP 2009050088W WO 2009088022 A1 WO2009088022 A1 WO 2009088022A1
Authority
WO
WIPO (PCT)
Prior art keywords
peptide
amino acid
acid sequence
cancer
seq
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2009/050088
Other languages
English (en)
Japanese (ja)
Inventor
Ikuro Maruyama
Teruto Hashiguchi
Shoji Natsugoe
Kenji Tanaka
Lyang-Ja Lee
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kagoshima University NUC
Protosera Inc
Original Assignee
Kagoshima University NUC
Protosera Inc
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 Kagoshima University NUC, Protosera Inc filed Critical Kagoshima University NUC
Priority to JP2009548938A priority Critical patent/JPWO2009088022A1/ja
Publication of WO2009088022A1 publication Critical patent/WO2009088022A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G01N33/57525—
    • 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
    • C07K14/745—Blood coagulation or fibrinolysis factors
    • C07K14/75—Fibrinogen
    • 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
    • C07K14/775—Apolipopeptides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/745—Assays involving non-enzymic blood coagulation factors
    • G01N2333/75—Fibrin; Fibrinogen
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/745—Assays involving non-enzymic blood coagulation factors
    • G01N2333/755—Factors VIII, e.g. factor VIII C [AHF], factor VIII Ag [VWF]

Definitions

  • the present invention relates to a novel diagnostic marker for various cancers including pancreatic cancer, and a diagnostic method for the disease using the same.
  • Pancreatic cancer has the lowest 5-year survival rate among common solid cancers, with approximately 20,000 and 30,000 deaths each year in Japan and the United States due to pancreatic cancer. Early detection is essential because pancreatic cancer is characterized by massive local invasion and early metastasis to the liver and lymph nodes, and surgical resection is the only credible cure. However, as often described as "silent organs", except for obstructive jaundice, the clinical symptoms of pancreatic cancer often do not become noticeable until the stage progresses, and the pancreas is located in the back of the abdomen Therefore, it is difficult to find pancreatic cancer with simple and inexpensive inspection means such as echo. As a result, only 20-40% of pancreatic cancer patients are surgically indicated. Therefore, development of a new test method that is inexpensive, simple, highly sensitive, and highly specific that enables early detection of pancreatic cancer is eagerly desired.
  • Non-Patent Document 1 analyzed a plasma sample of a pancreatic cancer patient using SELDI-TOFMS technology, and found four mass peaks (m / z7668,766) with significantly different peak intensities compared to healthy individuals. , 17,272, 28,080, 14,779) have been identified and reported as pancreatic cancer markers. However, it is unclear what protein these mass peaks originate from.
  • the fibrin monomer produced by cutting out FPA and FPB plays a main role in blood coagulation by polymerizing and insolubilizing.
  • the existence of a C-terminal fragment of fibrin ⁇ (positions 496-629 of the fibrinogen A ⁇ chain precursor polypeptide) has been reported, but its physiological significance has not been elucidated at all.
  • Apolipoprotein E is known as a causative gene for familial type III hyperlipidemia (E2) and a risk factor for Alzheimer's disease (E4).
  • E2 familial type III hyperlipidemia
  • E4 a risk factor for Alzheimer's disease
  • the relationship between apoE or its fragments and pancreatic cancer Is not reported at all.
  • An object of the present invention is to provide a novel diagnostic marker for pancreatic cancer and an effective diagnostic method for cancer using the same.
  • the present inventors examined serum collected from various cancer patients by mass spectrometry, and as a result, in pancreatic cancer patients, there was a marked increase compared to healthy individuals and cancer patients in other organs. Peptides having molecular weights of about 1616, about 2410 and about 2930 were found. As a result of analyzing the amino acid sequence, these peptides were found to be a peptide in which the Ser residue at position 3 of FPA was phosphorylated, a peptide consisting of a partial amino acid sequence at positions 170-192 of the apoE protein, and one of the fibrin ⁇ chain C-terminal fragment.
  • the present invention [1] In the subject, the amount of one or more peptides selected from the peptide group consisting of each amino acid sequence shown in SEQ ID NOs: 1, 2, and 3 in a biological sample collected from the subject is measured. Testing method for diagnosis of pancreatic cancer; [2] The method according to [1] above, wherein the peptide consisting of the amino acid sequence represented by SEQ ID NO: 1 is phosphorylated.
  • the method according to [5] The method according to any one of [1] to [4] above, comprising subjecting the biological sample to mass spectrometry; [6]
  • One or more antibodies selected from the group of antibodies specifically recognizing each peptide consisting of each amino acid sequence shown in SEQ ID NOs: 1, 2, and 3 are used.
  • pancreatic cancer can be determined quickly and accurately, so that early detection and early treatment of the disease are possible.
  • cancers in other organs can be diagnosed by appropriately combining the marker peptides of the present invention.
  • the present invention provides three new and useful diagnostic marker peptides for pancreatic cancer (hereinafter, the three peptides may be collectively referred to as “the peptide of the present invention”).
  • the first peptide of the present invention (also referred to as “peptide 1”) consists of the amino acid sequence shown in SEQ ID NO: 1.
  • the amino acid sequence corresponds to a fragment from the N-terminal Ala residue to the 16th Arg residue of the mature fibrinogen A ⁇ chain.
  • Peptide 1 is characterized in that the third Ser residue in the amino acid sequence is phosphorylated.
  • fibrinopeptide A FPA is produced by cleaving between the 16th Arg residue and the 17th Gly residue from the N-terminus of fibrinogen A ⁇ chain by thrombin.
  • Non-patent Document 2 the serum level of the peptide significantly decreases in the serum of prostate cancer, bladder cancer and breast cancer patients.
  • peptide 1 which is a phosphorylated FPA exhibits a very high value in the serum of pancreatic cancer patients, as opposed to the above cancer.
  • the serum level of peptide 1 is not as prominent as in pancreatic cancer patients, it is significantly increased in lung cancer, hematopoietic tumors, and gallbladder / bile duct cancer patients as compared with healthy individuals.
  • this peptide has a significantly lower serum level in prostate cancer and breast cancer patients than in healthy individuals, as well as endometrial cancer, colorectal cancer, esophagus. It is also significantly reduced in patients with cancer, cervical cancer and gastric cancer. Therefore, peptide 1 can also be used as a diagnostic marker for these cancers.
  • the second peptide of the present invention (also referred to as “peptide 2”) consists of the amino acid sequence shown in SEQ ID NO: 2.
  • the amino acid sequence corresponds to a fragment from the 170th Ala residue to the 192nd Leu residue from the N-terminus of apolipoprotein E (apoE).
  • the serum level of peptide 2 is significantly higher not only in patients with pancreatic cancer but also in patients with gallbladder / bile duct cancer as compared with cancer patients in healthy subjects and other organs.
  • peptide 2 can also be used as a diagnostic marker for these cancers.
  • the third peptide of the present invention (also referred to as “peptide 3”) consists of the amino acid sequence shown in SEQ ID NO: 3.
  • the amino acid sequence corresponds to a fragment from the 571th Ser residue to the 601st Tyr residue from the N-terminus of the fibrinogen A ⁇ chain precursor polypeptide.
  • Serum levels of peptide 3 are not as prominent as in pancreatic cancer patients, but cervical cancer, colorectal cancer, ovarian cancer, esophageal cancer, gallbladder / bile duct cancer, prostate cancer, endometrial cancer, stomach cancer, breast cancer and lung cancer In patients, it is significantly reduced compared to healthy individuals. Therefore, peptide 3 can also be used as a diagnostic marker for these cancers.
  • peptides 1 and 3 are fibrinogen A ⁇ chain fragments (degradation products), and peptide 2 is an apoE protein fragment. Therefore, the patient to be tested has a polymorphism or substitution comprising one or more amino acid substitutions, deletions, insertions or additions or combinations thereof within the partial amino acid sequence corresponding to these peptides of fibrinogen A ⁇ or apoE protein.
  • the amino acid sequence of the peptide to be detected should be understood as “the amino acid sequence having the polymorphism or allelic mutation in each of the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3.” It will be obvious to those skilled in the art.
  • the invention also provides a test for the diagnosis of pancreatic cancer in a patient by measuring the amount of one or more peptides of the invention in a biological sample from a patient suspected of suffering from pancreatic cancer.
  • test for diagnosis means measurement of the amount of the peptide and, if necessary, comparison with the measured value in a control sample.
  • the “patient suspected of having pancreatic cancer” may be subjectively suspected by the patient or based on some objective basis, but is preferably publicly known As a result of clinical examinations and / or examinations, patients who have been judged by a doctor to have a reasonable possibility of those diseases, or humans with an equivalent condition.
  • the peptide of the present invention is an intraepithelial cancer that is positioned at stage 0, which is the earliest of all six stages of disease classification (UICC, 1997). Even (Tis) is detected with a significant difference from that of a healthy person, and by adopting a periodic diagnosis using the peptide of the present invention as an index, there is a possibility that it can contribute to early detection and early treatment of pancreatic cancer.
  • the patient-derived biological sample to be the test sample is not particularly limited, but is preferably one that is less invasive to the patient, for example, blood, plasma, serum, saliva, urine, tears that can be easily collected from the living body And those collected relatively easily such as cerebrospinal fluid, bone marrow fluid, pleural effusion, ascites, joint fluid, aqueous humor, and vitreous humor.
  • serum or plasma it can be prepared by collecting blood from a patient according to a conventional method and separating the liquid component.
  • the peptide of the present invention as a detection target can be separated and removed in advance using a spin column or the like, if necessary, such as a high molecular weight protein fraction.
  • the detection of the peptide of the present invention in a biological sample can be performed, for example, by measuring various molecular weights of the biological sample, for example, gel electrophoresis or various separation and purification methods (eg, ion exchange chromatography, hydrophobic chromatography, affinity).
  • various separation and purification methods eg, ion exchange chromatography, hydrophobic chromatography, affinity.
  • ionization methods eg, electron impact ionization, field desorption, secondary ionization, fast atom bombardment, matrix-assisted laser desorption ionization (MALDI), electrospray ionization Etc.
  • mass spectrometers eg double-focusing mass spectrometer, quadrupole analyzer, time-of-flight mass spectrometer, Fourier transform mass spectrometer, ion cyclotron mass spectrometer, etc.
  • Detect bands, spots or peaks that match the molecular weight of the peptide Can be performed by the, not limited thereto.
  • the amino acid sequence of the peptide of the present invention is known, a method of preparing an antibody that recognizes the amino acid sequence and detecting the peptide by Western blotting or various immunoassays can be used more preferably. Furthermore, the hybrid detection method of the above method is also effective.
  • Peptides 1, 2 and 3 of the present invention have molecular weights (calculated values) of 1615.65, 2409.69 and 2930.28, respectively, but it goes without saying that the actual values may vary slightly depending on the measurement method / measurement equipment used. Absent. For example, in the case of a method using a mass spectrometer, it is preferable to measure the peak intensity appearing at the position of the calculated value ⁇ 0.5% (preferably ⁇ 0.3%, more preferably ⁇ 0.1%).
  • One of the particularly preferable measurement methods in the inspection method of the present invention is that a test sample is brought into contact with the surface of a plate used for time-of-flight mass spectrometry, and the mass of a component captured on the plate surface is measured using a time-of-flight mass spectrometer.
  • the method of measuring by is mentioned.
  • the plate that can be adapted to the time-of-flight mass spectrometer may be any plate as long as it has a surface structure that can efficiently adsorb the peptide of the present invention to be detected.
  • Such surface structures include, for example, functionalized glass, Si, Ge, GaAs, GaP, SiO 2 , SiN 4 , modified silicon, a wide range of gels or polymers (eg (poly) tetrafluoroethylene, (poly And vinylidene difluoride, polystyrene, polycarbonate, or combinations thereof).
  • functionalized glass Si, Ge, GaAs, GaP, SiO 2 , SiN 4
  • modified silicon eg (poly) tetrafluoroethylene, (poly And vinylidene difluoride, polystyrene, polycarbonate, or combinations thereof).
  • Examples of surface structures having a plurality of monomer or polymer sequences include, for example, linear and cyclic polymers of nucleic acids, polysaccharides, lipids, peptides having ⁇ -, ⁇ - or ⁇ -amino acids, gel surfaces used in chromatography Carriers (anionic / cationic compounds, hydrophobic compounds composed of carbon chains 1-18, hydrophilic compounds (eg, carriers cross-linked with silica, nitrocellulose, cellulose acetate, agarose, etc.), artificial homopolymers) (For example, polyurethane, polyester, polycarbonate, polyurea, polyamide, polyethyleneimine, polyarylene sulfide, polysiloxane, polyimide, polyacetate, etc.) Any known drug or natural compound is bound to any of the above compounds (covalent and non-covalent bonds). ) Coated with heteropolymer etc. Packaging and the like.
  • the support used as a plate for mass spectrometry is a substrate coated with a thin layer of polyvinylidene difluoride (PVDF), nitrocellulose or silica gel, particularly preferably PVDF [usually a plate for mass spectrometry
  • PVDF polyvinylidene difluoride
  • insulators glass, ceramics, plastics, resins, etc.
  • metals aluminum, stainless steel, etc.
  • conductive polymers composites thereof, etc.
  • an aluminum plate is preferably used, refer to WO 2004/031759.
  • the shape of the support can be appropriately devised into a shape suitable for the sample analyzer, in particular, the sample introduction port, but is not limited thereto.
  • a blot chip registered trademark
  • Protocera is preferably used as such a plate for mass spectrometry coated with a thin layer with PVDF.
  • the coating refers to a thin layer formed by depositing on a support in a state where coating molecules are dispersed, instead of overlaying a pre-formed structure like a membrane on the support.
  • the manner in which the coating molecules are deposited is not particularly limited, but means exemplified in a method for preparing a plate for mass spectrometry described later is preferably used.
  • the thickness of the thin layer can be appropriately selected within a range that does not adversely affect the transfer efficiency of the molecules contained in the tissue or cells and the measurement sensitivity of mass spectrometry, etc., for example, about 0.001 to about 100 ⁇ m, Preferably, it is about 0.01 to about 30 ⁇ m.
  • the plate for mass spectrometry (support) can be prepared by a method known per se, for example, the above preferred mass spectrometry plate is prepared by thinly coating the surface of the support with a coating molecule such as PVDF.
  • the Preferred examples of the coating means include application, spraying, vapor deposition, dipping, printing (printing), and sputtering.
  • the coating molecule is dissolved in an appropriate solvent, for example, an organic solvent such as dimethyl formamide (DMF) at an appropriate concentration (for example, about 1 to about 100 mg / mL) ( The coating molecule-containing solution) can be applied to the substrate using a suitable tool such as a brush.
  • an appropriate solvent for example, an organic solvent such as dimethyl formamide (DMF) at an appropriate concentration (for example, about 1 to about 100 mg / mL)
  • DMF dimethyl formamide
  • the coating molecule-containing solution can be applied to the substrate using a suitable tool such as a brush.
  • the coating molecule-containing solution prepared in the same manner as described above may be put in a sprayer and sprayed so that PVDF is uniformly deposited on the substrate.
  • the coating molecule (which may be solid or solution) is heated and vaporized in a vacuum tank containing the substrate using a normal vacuum deposition apparatus for producing an organic thin film. A thin layer of molecules can be formed.
  • immersion the substrate may be immersed in a coating molecule-containing solution prepared in the same manner as described above.
  • various printing techniques that can be normally used depending on the material of the substrate can be appropriately selected and used. For example, screen printing or the like is preferably used.
  • sputtering for example, a DC high voltage is applied between the substrate and the coating molecule while introducing an inert gas (eg, Ar gas) in a vacuum, and the ionized gas is caused to collide with the molecule.
  • an inert gas eg, Ar gas
  • the repelled coating molecules can be deposited on the substrate to form a thin layer.
  • the coating may be applied to the entire surface of the substrate, or may be applied only to the surface (fraction) subjected to mass spectrometry.
  • the coating molecule can be used in a suitable form depending on the coating means.
  • the coating molecule can be applied to the substrate in the form of a coating molecule-containing solution, a coating molecule-containing vapor, a solid coating molecule, etc. It is preferable to apply in the form. “Apply” refers to bringing a coating molecule into contact with the support so that the coating molecules remain and deposit on the support after contact.
  • the amount of application is not particularly limited, and examples of the coating molecular weight include about 10 to about 100,000 ⁇ g / cm 2 , preferably about 50 to about 5,000 ⁇ g / cm 2 . After the application, the solvent is removed by natural drying, vacuum drying or the like.
  • the surface of the substrate in the mass spectrometry plate may be modified (processed) in advance by an appropriate physical or chemical technique before coating with a coating molecule. Specifically, techniques such as polishing the plate surface, scratching, acid treatment, alkali treatment, glass treatment (tetramethoxysilane, etc.) are exemplified.
  • Transfer of the test sample to the mass spectrometric plate (support) can be performed by leaving the patient-derived biological sample as the test sample untreated or after removing and concentrating the high molecular weight protein using an antibody column or other method.
  • -It is performed by subjecting to polyacrylamide gel electrophoresis or isoelectric focusing, and transferring (blotting) the gel after contact with the plate.
  • a known transfer device can be used.
  • the transfer method itself is known.
  • electrotransfer is used.
  • the sample developed on the gel after the electrophoresis is transferred to the plate for mass spectrometry by various methods (diffusion, electric force, etc.).
  • As a buffer used for electrotransfer it is preferable to use a buffer having a pH of 7 to 9 and a low salt concentration.
  • Tris buffer Tris buffer, phosphate buffer, borate buffer, and acetate buffer.
  • the buffer include sodium borate-hydrochloric acid buffer, tris-borate / EDTA, borate / ACN, and the like.
  • the acetate buffer include tris-acetate / EDTA. Preferred are tris / glycine / methanol buffer and sodium borate-hydrochloric acid buffer.
  • compositions of the tris / glycine / methanol buffer include Tris 10-15 ⁇ mM, glycine 70-120 ⁇ mM, and methanol 7-13%.
  • An example of the composition of the sodium borate-hydrochloric acid buffer is about 5 to 20 mM sodium borate.
  • the molecules present in the test sample including the target molecules are efficiently captured on the surface of the support.
  • a reagent called matrix is added to absorb the laser light and promote ionization of analyte molecules through energy transfer, which is advantageous for later mass spectrometry (when using MALDI method) You can also.
  • the matrix those known in mass spectrometry can be used.
  • IAA indoleacrylic acid
  • DVB 2,5-dihydroxybenzoic acid
  • CHCA ⁇ -cyano-4-hydroxycinammic acid
  • it is DHB or CHCA.
  • the presence and amount of the peptide of the present invention can be identified from the information on the molecular weight by mass spectrometry of molecules in the test sample captured on the support surface by the above method.
  • the mass spectrometer measures the molecular weight of a substance by ionizing a gaseous sample and then putting the molecule or molecular fragment into an electromagnetic field, separating it by mass number / charge number from its movement, and obtaining the spectrum of the substance.
  • -It is a device to detect.
  • MALDI Matrix-assisted laser deionization
  • a sample and a matrix that absorbs laser light are mixed, dried and crystallized, and ionized analytes are brought into vacuum by ionization by energy transfer from the matrix and instantaneous heating by laser irradiation.
  • MALDI-TOFMS method which uses time-of-flight mass spectrometry (TOFMS), which analyzes the mass number based on the time-of-flight difference of sample molecular ions by initial acceleration.
  • TOFMS time-of-flight mass spectrometry
  • the presence or absence and the amount of the target molecule in the test sample can be identified based on the molecular weight information of the target molecule.
  • information from the mass spectrometer as differential information by comparing it with mass spectrometry data in a biological sample derived from a healthy person using an arbitrary program. It will be appreciated that such programs are well known and those skilled in the art can easily construct or modify such programs using known information processing techniques.
  • each of the above steps is performed using a blot chip manufactured by Protocera as a plate for mass spectrometry, and the peptides of the present invention are quantitatively compared (differential analysis) using a MALDI mass spectrometer. Furthermore, if necessary, the peptides remaining on the same chip can be identified. Alternatively, up to quantitative comparison (differential analysis) of test samples is performed using a blot chip system manufactured by Protocera, and the peptide is identified by a combination device (LC-MS / LC / MS / MS). MS).
  • the measurement of the peptide of the present invention in the test method of the present invention can also be performed using an antibody against it.
  • an optimized immunoassay system is constructed and a kit is made, the peptide can be detected with high sensitivity and high accuracy without using a special device such as the mass spectrometer. It is particularly useful in that it can.
  • the antibody against the peptide of the present invention can be prepared, for example, by isolating and purifying the peptide of the present invention from a biological sample derived from a patient expressing the peptide and immunizing an animal using the peptide as an antigen.
  • the peptide is partially digested with peptidase or the like, the amino acid sequence of the obtained fragment is determined by the Edman method or the like, and the nucleic acid encoding the peptide based on the sequence
  • a hybridizable oligonucleotide is synthesized and a cDNA library derived from the patient is used as a template to obtain a cDNA encoding a protein containing the peptide, or a cDNA derived from the patient is used as a primer.
  • the peptide of the present invention can be prepared in large quantities by collecting the recombinant peptide. You can.
  • the peptide of the present invention can be obtained by using a cell-free transcription / translation system using the cDNA obtained as described above as a template. Further, it can be prepared in a large amount by an organic synthesis method.
  • the peptides 1, 2 and 3 of the present invention are peptides consisting of the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively. Therefore, the antibody against the peptide of the present invention can be synthesized, for example, by synthesizing all or part of the amino acid sequence using a known peptide synthesis method based on the amino acid sequence information, or by fibrinogen A ⁇ isolated by a conventional method. It is desirable to cleave the chain or apoE protein with an appropriate peptidase or the like to obtain a peptide fragment containing all or part of the sequence of the peptide of the present invention, and prepare this as an immunogen.
  • the antibody against the marker peptide of the present invention may be either a polyclonal antibody or a monoclonal antibody, and can be prepared by a known immunological technique.
  • the antibody includes not only a complete antibody molecule but also a fragment thereof, and examples thereof include Fab, F (ab ′) 2, ScFv, and minibody.
  • the polyclonal antibody is cross-linked to a carrier protein such as bovine serum albumin or KLH (Keyhole Limpet Hemocyanin) prepared by any of the above methods or other methods, or a partial peptide thereof.
  • a carrier protein such as bovine serum albumin or KLH (Keyhole Limpet Hemocyanin) prepared by any of the above methods or other methods, or a partial peptide thereof.
  • the antibody titer of the collected serum is measured by a known antigen-antibody reaction, and the increase is confirmed), and it can be obtained by collecting the whole blood about 3 to about 10 days after the final immunization and purifying the antiserum.
  • animals to which the antigen is administered include mammals such as rats, mice, rabbits, goats, guinea pigs, and hamsters
  • Monoclonal antibodies can be obtained by cell fusion methods (for example, Takeshi Watanabe, Principles of Cell Fusion Methods and Production of Monoclonal Antibodies, Akira Taniuchi, Toshitada Takahashi, “Monoclonal Antibodies and Cancer: Basic and Clinical”, 2-14). Page, Science Forum Publishing, 1985).
  • the peptide of the present invention or a partial peptide thereof is administered to a mouse subcutaneously or intraperitoneally 2-4 times together with a commercially available adjuvant, and the spleen or lymph node is collected about 3 days after the final administration, and white blood cells are collected.
  • the leukocytes and myeloma cells are cell-fused to obtain a hybridoma that produces a monoclonal antibody against the peptide.
  • the cell fusion may be PEG method [J. Immunol. Methods, 81 (2): 223-228 (1985)] or voltage pulse method [Hybridoma, 7 (6): 627-633 (1988)].
  • a hybridoma producing a desired monoclonal antibody can be selected by detecting an antibody that specifically binds to an antigen from the culture supernatant using a known EIA or RIA method or the like.
  • the culture of the hybridoma producing the monoclonal antibody can be performed in vitro or in vivo such as mouse or rat, or preferably mouse ascites, and the antibody can be obtained from the culture supernatant of the hybridoma and the ascites of the animal, respectively. .
  • non-phosphorylated FPA and fragments thereof may also be present in the test sample. These peptide groups do not show significant variation in pancreatic cancer, but any of these may be significantly higher in other diseases. If the antibody against peptide 1 of the present invention has cross-reactivity with one or more other N-terminal fragments of fibrinogen A ⁇ chain, the possibility of misdiagnosing other diseases as pancreatic cancer (false positive) is increased. Therefore, the antibody against peptide 1 of the present invention used in the test method of the present invention is preferably a highly specific antibody that does not cross-react with the N-terminal fragment of fibrinogen A ⁇ chain other than the peptide.
  • Such an antibody can be obtained by reacting a plurality of monoclonal antibodies obtained as described above with the other fragments and selecting an antibody that does not cross-react with them.
  • peptide 1 of the present invention is phosphorylated FPA, it is desirable that an antibody against peptide 1 is capable of specifically recognizing the phosphorylated site.
  • the test method of the present invention using the antibody of the present invention is not particularly limited, and the amount of antibody, antigen or antibody-antigen complex corresponding to the amount of antigen in the test sample is determined by chemical or physical means. Any measurement method may be used as long as it is a measurement method that is detected and calculated from a standard curve prepared using a standard solution containing a known amount of antigen. For example, nephrometry, competition method, immunometric method and sandwich method are preferably used.
  • a labeling agent used in a measurement method using a labeling substance for example, a radioisotope, an enzyme, a fluorescent substance, a luminescent substance, or the like is used.
  • the radioisotope for example, [ 125 I], [ 131 I], [ 3 H], [ 14 C] and the like are used.
  • the enzyme is preferably stable and has a large specific activity.
  • ⁇ -galactosidase, ⁇ -glucosidase, alkaline phosphatase, peroxidase, malate dehydrogenase and the like are used.
  • the fluorescent substance for example, fluorescamine, fluorescein isothiocyanate and the like are used.
  • luminescent substance for example, luminol, luminol derivatives, luciferin, lucigenin and the like are used.
  • a biotin-avidin system can also be used for binding of an antibody or antigen and a labeling agent.
  • the carrier include insoluble polysaccharides such as agarose, dextran, and cellulose, synthetic resins such as polystyrene, polyacrylamide, and silicon, or glass.
  • a test sample is reacted with an insolubilized antibody of the present invention (primary reaction), and another labeled antibody of the present invention is reacted (secondary reaction), followed by a labeling agent on an insolubilized carrier.
  • primary reaction an insolubilized antibody of the present invention
  • secondary reaction another labeled antibody of the present invention
  • a labeling agent on an insolubilized carrier By measuring the amount (activity), the amount of the peptide of the present invention in the test sample can be quantified.
  • the primary reaction and the secondary reaction may be performed in the reverse order, or may be performed simultaneously or at different times.
  • the monoclonal antibody against the polypeptide of the present invention can also be used in measurement systems other than the sandwich method, such as a competitive method, an immunometric method, or nephrometry.
  • a competitive method the antigen in the test sample and the labeled antigen are reacted competitively with the antibody, and then the unreacted labeled antigen (F) and the labeled antigen (B) bound to the antibody are separated ( B / F separation), measure the amount of labeled B or F, and quantify the amount of antigen in the test sample.
  • a soluble antibody is used as an antibody
  • a B / F separation is performed using polyethylene glycol
  • a liquid phase method using a second antibody against the antibody and a solid-phased antibody is used as the first antibody, or
  • the first antibody is soluble
  • the second antibody is a solid phase method using a solid phase antibody.
  • the antigen of the test sample and the immobilized antigen are competitively reacted with a certain amount of labeled antibody, and then the solid phase and the liquid phase are separated, or the antigen and excess in the test sample are separated.
  • the solid phase antigen is added, and then the solid phase antigen is added to bind the unreacted labeled antibody to the solid phase, and then the solid phase and the liquid phase are separated.
  • the amount of label in any phase is measured to quantify the amount of antigen in the test sample.
  • the amount of insoluble precipitate produced as a result of the antigen-antibody reaction in a gel or solution is measured. Laser nephrometry using laser scattering is preferably used even when the amount of antigen in the test sample is small and only a small amount of precipitate is obtained.
  • peptides of the present invention are composed of proteolytic products, various molecules such as undegraded proteins and similar peptides with a common cleavage site may affect the measured value in the usual “sandwich ELISA system”. Therefore, in the first step, a biological sample is immunoaffinity purified with an antibody, the fraction bound to the antibody is subjected to mass spectrometry in the second step, and so-called immunomass spectrometry, in which identification and quantification are performed based on a precise molecular weight. (For example, see Rapid Commun. Mass Spectrom. 2007, 21: 352-358).
  • a biomarker peak is not observed even if the sample is directly measured with a MALDI mass spectrometer.
  • undegraded protein is also a similar peptide. Is completely separated by a mass spectrometer, and can be quantified with high specificity and sensitivity based on the exact molecular weight of the biomarker.
  • the antibody is immobilized on the surface of a probe that can be adapted to a mass spectrometer as described above, and a test sample is applied to the antibody on the probe.
  • a test sample is applied to the antibody on the probe. Examples include a method of detecting a peak corresponding to the molecular weight of a marker peptide recognized by the antibody by subjecting the biological sample component captured by the antibody to mass spectrometry.
  • the level of peptide 1 in a sample derived from a subject measured by any of the above methods is about 2 times or more, preferably about 5 times or more than the level of peptide 1 in a control sample from a healthy person
  • the subject can be diagnosed as having a high possibility of having pancreatic cancer.
  • the value is statistically significantly higher than the peptide level in the control sample.
  • it can be diagnosed that the subject is likely to have pancreatic cancer.
  • the level of peptide 1 in a sample derived from a subject is statistically significantly higher than the level of peptide 1 in a control sample derived from a healthy subject, Can be diagnosed as having a high possibility of having lung cancer, hematopoietic tumor or gallbladder / bile duct cancer in addition to pancreatic cancer.
  • the level of peptide 1 in a sample from a subject is statistically significantly lower than the level of peptide 1 in a control sample from a healthy person, the subject is prostate cancer, It can be diagnosed that there is a high possibility of having breast cancer, endometrial cancer, colorectal cancer, esophageal cancer, cervical cancer or stomach cancer.
  • the level of peptide 2 in a sample derived from a subject measured by any of the above methods is about 2 times or more, preferably about 5 times or more than the level of peptide 2 in a control sample from a healthy person
  • the subject can be diagnosed as having a high possibility of having pancreatic cancer or gallbladder / bile duct cancer.
  • the value was statistically significantly higher than the peptide level in the control sample.
  • pancreatic cancer can be differentiated from gallbladder / bile duct cancer by combining peptide 1 and / or peptide 3 as a marker.
  • the level of peptide 2 in the sample derived from the subject is statistically significantly higher than the level of peptide 2 in the control sample derived from the healthy subject, Can be diagnosed as having a high possibility of having ovarian cancer, esophageal cancer or hematopoietic tumor in addition to pancreatic cancer and gallbladder / bile duct cancer.
  • the subject can be diagnosed as having a high probability of having pancreatic cancer.
  • the level of peptide 3 in the sample derived from the subject is statistically significantly lower than the level of peptide 3 in the control sample derived from the healthy subject, the subject is diagnosed with cervical cancer. It can be diagnosed that there is a high possibility of suffering from colon / rectal cancer, ovarian cancer, esophageal cancer, gallbladder / bile duct cancer, prostate cancer, endometrial cancer, stomach cancer, breast cancer or lung cancer.
  • the test method of the present invention is preferably carried out by collecting biological samples from patients in time series and examining the time course of the expression of the peptide of the present invention in each sample.
  • the collection interval of the biological sample is not particularly limited, but it is desirable to sample as frequently as possible within a range that does not impair the patient's QOL.
  • plasma or serum is used as a sample, blood is collected at intervals of about 1 minute to about 12 hours It is preferable to carry out.
  • peptides 1 and 3 tend to increase in serum level as the stage of pancreatic cancer progresses, at least in stage 0-II of the disease classification.
  • Peptide 2 shows a markedly high value at stage 0, and maintains a significantly high value as compared with healthy individuals even when the stage progresses. Therefore, when the level of the peptide of the present invention decreases with time, it can be determined that there is a high possibility that the pathological condition of pancreatic cancer in the patient is improved.
  • the method for examining pancreatic cancer based on the above time-series sampling is such that when a treatment measure for the disease is taken for a patient who is a subject between the previous sampling and the current sampling, treatment by the measure is performed. It can be used to evaluate the effect. That is, for a sample sampled before and after treatment, when it is determined that the condition after treatment is improved compared to the condition before treatment, it can be evaluated that the treatment is effective. On the other hand, when it is determined that the condition after treatment is not improved or further deteriorated as compared with the condition before treatment, it can be evaluated that the treatment has no effect.
  • the peptide of the present invention can also provide an active drug discovery target for pancreatic cancer in addition to diagnosis. That is, when the peptide itself has a physiological function in the direction of treatment (remission) of the disease (referred to as “therapeutic peptide”), by administering to the patient a substance that increases the amount or activity of the peptide, When the peptide itself has physiological functions in the direction of exacerbation of the disease (referred to as “exacerbation peptide”), the disease can be treated by administering a substance that reduces the amount or activity of the peptide.
  • therapeutic peptide a physiological function in the direction of treatment (remission) of the disease
  • exacerbation peptide the disease can be treated by administering a substance that reduces the amount or activity of the peptide.
  • the invention also increases the amount or activity of the peptide when the peptide of the invention acts as a therapeutic peptide, and / or the amount or amount of the peptide when the peptide of the invention acts as an exacerbation peptide.
  • Methods of treating pancreatic cancer by reducing activity are provided.
  • the therapeutic method specifically comprises an effective amount of a substance that increases the amount or activity of the peptide of the present invention as a therapeutic peptide and / or a substance that decreases the amount or activity of the peptide of the present invention as an exacerbation peptide, Administration to patients with pancreatic cancer.
  • the present invention also includes a substance that increases the amount or activity of the peptide of the present invention as a therapeutic peptide and / or a substance that decreases the amount or activity of the peptide of the present invention as an exacerbation peptide.
  • a therapeutic agent examples include the peptide itself or a molecule having an agonistic action similar thereto.
  • a non-neutralizing antibody, preferably an agonist antibody, of the peptide can also be mentioned.
  • examples of the substance that reduces the activity of the peptide of the present invention as an exacerbation peptide include a molecule having an antagonistic action of the peptide, or a neutralizing antibody against the peptide.
  • the substance that increases the production of the peptide of the present invention as a therapeutic peptide includes a degrading enzyme that releases the peptide from a parent protein (fibrinogen, apoE) existing in the living body, the N-terminal side of the peptide and / or C Further comprising an amino acid sequence that is recognized and cleaved by the decomposing enzyme on the terminal side, a substrate or a substrate analog molecule of the decomposing enzyme, a molecule (including an analogous compound) that promotes the production of the decomposing enzyme, Examples include molecules that promote, molecules that suppress the production of inhibitors of the degrading enzyme, and the like.
  • a substrate or substrate analog molecule of a decomposing enzyme thus identified that is, a peptide molecule further comprising an amino acid sequence recognized and cleaved by the decomposing enzyme on the N-terminal side and / or C-terminal side of the peptide, Since it is cleaved by the degrading enzyme in the body to release the peptide of the present invention or its analog molecule as a therapeutic peptide, the same therapeutic effect can be obtained.
  • substances that promote the production and / or activity of the identified degrading enzymes can also indirectly increase the production of the peptides of the invention as therapeutic peptides. If the target degrading enzyme is identified, these substances can be screened or molecularly designed by a method known per se.
  • the substance that reduces the production of the peptide of the present invention as an exacerbation peptide includes a molecule that suppresses the production of a degrading enzyme that liberates the peptide from a protein present in the living body, an inhibitor of the decomposing enzyme, and the production of the inhibitor And molecules that promote
  • the degrading enzyme that liberates the peptide of the present invention as an exacerbation peptide can be searched and identified by the same method as the peptide of the present invention as the therapeutic peptide.
  • a substance that suppresses (inhibits) the production or activity of the degrading enzyme directly or indirectly can be screened or molecularly designed by a method known per se.
  • compositions for oral administration include solid or liquid dosage forms, specifically tablets (including dragees and film-coated tablets), pills, granules, powders, capsules (including soft capsules). Syrup, emulsion, suspension and the like.
  • Such a composition is produced by a method known per se, and contains a carrier, diluent or excipient usually used in the pharmaceutical field.
  • lactose, starch, sucrose, magnesium stearate and the like are used as carriers and excipients for tablets.
  • a composition for parenteral administration for example, injections, suppositories and the like are used, and injections are intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, intravenous injections, intraarticular injections. Includes dosage forms such as agents.
  • Such an injection is prepared according to a method known per se, for example, by dissolving, suspending or emulsifying the above compound or a salt thereof in a sterile aqueous or oily liquid usually used for injection.
  • aqueous solution for injection for example, isotonic solutions containing physiological saline, glucose and other adjuvants are used, and suitable solubilizers such as alcohol (eg, ethanol), polyalcohol (eg, Propylene glycol, polyethylene glycol), nonionic surfactants (eg, polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)) and the like may be used in combination.
  • alcohol eg, ethanol
  • polyalcohol eg, Propylene glycol, polyethylene glycol
  • nonionic surfactants eg, polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)
  • oily liquid for example, sesame oil, soybean oil and the like are used, and benzyl benzoate, benzyl alcohol and the like may be used in combination as a solubilizing agent.
  • the prepared injection solution is usually filled in a suitable amp
  • compositions are conveniently prepared in dosage unit form to suit the dosage of the active ingredient.
  • dosage form of such a dosage unit include tablets, pills, capsules, injections (ampoules), suppositories, etc., and usually 5 to 500 mg, particularly 5 to 100 mg for injections, Other dosage forms preferably contain 10 to 250 mg of the above compound.
  • Each of the above-described compositions may be mixed with a substance that increases the amount or activity of the peptide of the present invention as the therapeutic peptide or a substance that decreases the amount or activity of the peptide of the present invention as an exacerbation peptide.
  • Other active ingredients may be contained as long as the action is not caused.
  • the preparation thus obtained is safe and has low toxicity, it can be administered, for example, orally or parenterally to humans.
  • a substance that increases the amount or activity of the peptide of the present invention as a therapeutic peptide and a substance that decreases the amount or activity of the peptide of the present invention as an exacerbation peptide are determined by its action, administration route, patient severity, although there are differences depending on age, body weight, drug acceptability, etc., for example, the amount of active ingredient per day for an adult is about 0.0008 to about 25 mg / kg, preferably about 0.008 to about 2 mg / kg, This can be administered once or in several divided doses.
  • Example 1 Profiling analysis using BlotChip 1.5 ⁇ L of serum of various cancer patients and healthy subject serum were mixed with 4.5 ⁇ L of sample treatment solution for electrophoresis (NuPAGE (registered trademark) LDS Sample Buffer 4x; Invitrogen) for 10 minutes at 70 ° C. After heat treatment, it was applied to a 4-12% gradient polyacrylamide gel (Invitorogen) and subjected to electrophoresis. After the completion of electrophoresis, the gel was cut out, layered on BLOTCHIP (registered trademark) (Protosera, Inc.), and transferred in an electric transfer buffer (BLOTBuffer TM ; Protosera, Inc.) at 90 mA for 120 minutes.
  • BLOTCHIP registered trademark
  • BLOTBuffer TM Electric transfer buffer
  • Example 2 Identification of peptides by de novo MS / MS analysis on BlotChip
  • matrix-assisted laser desorption ionization Using time-of-flight (MALDI-TOF) mass spectrometer (Ultra-FlexII by Bruker Daltonics), Bradykinin, AngiotensinII, AngiotensinI, SubstanceP, Bombesin, Renin Substrate, ACTH Clip ⁇ 1-17 ⁇ , ACTH Clip ⁇ 18 -39 ⁇ , mass calibration was performed using Somatostatin.
  • MALDI-TOF time-of-flight
  • peptides 1 to 3 were identified as peptides each having the amino acid sequence shown in SEQ ID NOs: 1 to 3, respectively.
  • peptide 1 was phosphorylated from FPA which is a thrombin degradation product of fibrinogen
  • peptide 2 was a fragment corresponding to positions 170-192 of apoE protein
  • peptide 3 was 571 of fibrinogen
  • a ⁇ chain precursor polypeptide -It became clear that it was a fragment corresponding to position 601.
  • the clinical test method using the novel diagnostic marker for pancreatic cancer according to the present invention is useful in that pancreatic cancer can be determined quickly and accurately, so that early detection and early treatment of the disease are possible.
  • the peptide of the present invention as a measurement target in the present invention can itself be a drug discovery target in these diseases, it can be used for screening for novel therapeutic agents for pancreatic cancer and for treating the diseases using them. This is extremely useful.
  • This application is based on Japanese Patent Application No. 2008-000776 filed in Japan, the contents of which are incorporated in full herein.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Hematology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention porte sur un procédé de test pour diagnostiquer un cancer du pancréas chez un sujet, lequel procédé est caractérisé par la mesure de la quantité d'au moins un peptide choisi dans le groupe constitué par les peptides comprenant les séquences d'acides aminés illustrées dans la séquence ID n° : 1, 2 et 3, respectivement, dans un échantillon biologique (de préférence, un fluide corporel, idéalement un sérum, un plasma ou un fluide semblable) collecté à partir du sujet (de préférence par spectrométrie de masse ou à l'aide d'un anticorps capable de reconnaître de façon spécifique le peptide).
PCT/JP2009/050088 2008-01-07 2009-01-07 Nouveau marqueur de cancer et diagnostic l'utilisant Ceased WO2009088022A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2009548938A JPWO2009088022A1 (ja) 2008-01-07 2009-01-07 新規癌マーカーおよびそれを用いた診断

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2008000776 2008-01-07
JP2008-000776 2008-01-07

Publications (1)

Publication Number Publication Date
WO2009088022A1 true WO2009088022A1 (fr) 2009-07-16

Family

ID=40853134

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2009/050088 Ceased WO2009088022A1 (fr) 2008-01-07 2009-01-07 Nouveau marqueur de cancer et diagnostic l'utilisant

Country Status (2)

Country Link
JP (1) JPWO2009088022A1 (fr)
WO (1) WO2009088022A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011052380A1 (fr) * 2009-10-28 2011-05-05 日東紡績株式会社 PROCÉDÉ D'IMMUNOESSAI DE PEPTIDE DE 5,9 kDa
JP2011099858A (ja) * 2009-11-06 2011-05-19 Chiba Univ 胆道がんの新規バイオマーカー
JP2012051822A (ja) * 2010-08-31 2012-03-15 Institute Of Physical & Chemical Research 肺癌診断用ポリペプチド、肺癌の検出方法、および治療効果の評価方法
JP2013506142A (ja) * 2009-10-01 2013-02-21 フェノメノーム ディスカバリーズ インク 膵臓癌の血清ベースのバイオマーカー並びに疾患検出及び診断のためのその使用
CN109425739A (zh) * 2017-08-31 2019-03-05 复旦大学 一组蛋白作为肿瘤标志物在制备恶性肿瘤诊断试剂和试剂盒中的用途
JPWO2021221179A1 (fr) * 2020-04-28 2021-11-04

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112255334B (zh) * 2020-09-28 2022-06-17 复旦大学 用于区分交界性和恶性卵巢肿瘤的小分子标志物及其应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006308533A (ja) * 2005-05-02 2006-11-09 Mcbi:Kk 新規肝がんバイオマーカーおよび該バイオマーカーを用いた肝がんの検出方法
WO2006125021A2 (fr) * 2005-05-16 2006-11-23 Dana Farber Cancer Institute, Inc. Methodes de detection du cancer
WO2006129717A1 (fr) * 2005-05-31 2006-12-07 National University Corporation Chiba University Marqueur tumoral pour la detection de cancer pancreatique et kit de detection de cancer pancreatique l’employant
WO2008072676A1 (fr) * 2006-12-12 2008-06-19 Kagoshima University Marqueur innovant d'affection et diagnostic a l'aide de celui-ci
JP2009029731A (ja) * 2007-07-25 2009-02-12 Japan Health Science Foundation 膵癌の新規腫瘍マーカー

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006308533A (ja) * 2005-05-02 2006-11-09 Mcbi:Kk 新規肝がんバイオマーカーおよび該バイオマーカーを用いた肝がんの検出方法
WO2006125021A2 (fr) * 2005-05-16 2006-11-23 Dana Farber Cancer Institute, Inc. Methodes de detection du cancer
WO2006129717A1 (fr) * 2005-05-31 2006-12-07 National University Corporation Chiba University Marqueur tumoral pour la detection de cancer pancreatique et kit de detection de cancer pancreatique l’employant
WO2008072676A1 (fr) * 2006-12-12 2008-06-19 Kagoshima University Marqueur innovant d'affection et diagnostic a l'aide de celui-ci
JP2009029731A (ja) * 2007-07-25 2009-02-12 Japan Health Science Foundation 膵癌の新規腫瘍マーカー

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
VILLANUEVA J. ET AL.: "Differential exoprotease activities confer tumor-specific serum peptidome patterns", THE JOURNAL OF CLINICAL INVESTIGATION, vol. 116, no. 1, January 2006 (2006-01-01), pages 271 - 284 *

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014240841A (ja) * 2009-10-01 2014-12-25 フェノメノーム ディスカバリーズ インク 膵臓癌の血清ベースのバイオマーカー並びに疾患検出及び診断のためのその使用
US11079385B2 (en) 2009-10-01 2021-08-03 Med-Life Discoveries Lp Serum-based biomarkers of pancreatic cancer and uses thereof for disease detection and diagnosis
US10656155B2 (en) 2009-10-01 2020-05-19 Med-Life Discoveries Lp Serum-based biomarkers of pancreatic cancer and uses thereof for disease detection and diagnosis
US10024857B2 (en) 2009-10-01 2018-07-17 Med-Life Discoveries Lp Serum-based biomarkers of pancreatic cancer and uses thereof for disease detection and diagnosis
JP2013506142A (ja) * 2009-10-01 2013-02-21 フェノメノーム ディスカバリーズ インク 膵臓癌の血清ベースのバイオマーカー並びに疾患検出及び診断のためのその使用
WO2011052380A1 (fr) * 2009-10-28 2011-05-05 日東紡績株式会社 PROCÉDÉ D'IMMUNOESSAI DE PEPTIDE DE 5,9 kDa
CN102597772B (zh) * 2009-10-28 2014-08-20 日东纺绩株式会社 5.9kDa肽的免疫学测定方法
JP5585587B2 (ja) * 2009-10-28 2014-09-10 日東紡績株式会社 ５．９ｋＤａペプチドの免疫学的測定方法
CN102597772A (zh) * 2009-10-28 2012-07-18 日东纺绩株式会社 5.9kDa肽的免疫学测定方法
US9017959B2 (en) 2009-10-28 2015-04-28 Nitto Boseki Co., Ltd. 5.9 kDa peptide immunoassay method
EP2495564A4 (fr) * 2009-10-28 2013-07-10 Nitto Boseki Co Ltd PROCÉDÉ D'IMMUNOESSAI DE PEPTIDE DE 5,9 kDa
JP2011099858A (ja) * 2009-11-06 2011-05-19 Chiba Univ 胆道がんの新規バイオマーカー
JP2012051822A (ja) * 2010-08-31 2012-03-15 Institute Of Physical & Chemical Research 肺癌診断用ポリペプチド、肺癌の検出方法、および治療効果の評価方法
CN109425739B (zh) * 2017-08-31 2022-03-18 复旦大学 一组蛋白作为肿瘤标志物在制备恶性肿瘤诊断试剂和试剂盒中的用途
CN109425739A (zh) * 2017-08-31 2019-03-05 复旦大学 一组蛋白作为肿瘤标志物在制备恶性肿瘤诊断试剂和试剂盒中的用途
JPWO2021221179A1 (fr) * 2020-04-28 2021-11-04
WO2021221179A1 (fr) * 2020-04-28 2021-11-04 国立大学法人高知大学 Établissement d'un modèle de souris à l'aide d'un organoïde du cancer du pancréas humain
JP7702110B2 (ja) 2020-04-28 2025-07-03 国立大学法人高知大学 ヒト膵癌オルガノイドを用いたマウスモデルの樹立

Also Published As

Publication number Publication date
JPWO2009088022A1 (ja) 2011-05-26

Similar Documents

Publication Publication Date Title
JPWO2008072676A1 (ja) 新規疾患マーカーおよびそれを用いた診断
JP5424331B2 (ja) 肝疾患診断用バイオマーカー
US20100240088A1 (en) Peptide Markers for Diagnosis of Angiogenesis
EP2548888B1 (fr) Peptide spécifique d'une pathologie parodontale et traitement et diagnostic d'une pathologie parodontale l'incluant
WO2009088022A1 (fr) Nouveau marqueur de cancer et diagnostic l'utilisant
US7297556B2 (en) Method of diagnosing nephrotic syndrome
JP5714285B2 (ja) 妊娠高血圧症候群マーカーおよびそれを用いた診断
JP2010071953A (ja) 乳癌マーカーおよびそれを用いた診断
JP5410997B2 (ja) うつ病およびうつ状態のマーカーおよびそれを用いた検出・診断
JP2011117813A (ja) 変形性関節症のマーカーおよびそれを用いた診断
JP2011080775A (ja) 妊娠高血圧症候群マーカーおよびそれを用いた診断
JP2014020941A (ja) 大腸癌のマーカーおよびそれを用いた診断
JP2015108515A (ja) 大腸癌の診断のための検査方法
JP6521306B2 (ja) 飲酒に伴う高血圧発症の予測用ペプチドおよびそれを用いた飲酒に伴う高血圧発症の予測方法
JP2007332120A (ja) 新規病態マーカー及びそれを用いた病態診断
JP7300660B2 (ja) 大腸がんの検出方法
JP2014025868A (ja) がん転移マーカーおよびそれを用いた診断
JP7336097B2 (ja) 乳がんに関するペプチドマーカー
JP5117336B2 (ja) 多発性硬化症またはｎｍｏの検査マーカーの測定方法
KR20130102693A (ko) 유방암 진단용 단백질 마커 아포리포단백질 （ａ）, 이의 검출 방법 및 이에 대한 항체를 포함하는 유방암 진단키트
US20080089889A1 (en) Novel Bone Metastasis Marker Peptide and Method of Diagnosing Bone Metastasis by Using the Same
JP2024117521A (ja) 膵臓がんの検出方法
KR100559374B1 (ko) 종양 진단 마커로서의 헤모글로빈
JP2007314493A (ja) モノクローナル抗体、その製造方法、及び用途
JP5924659B2 (ja) 免疫複合体の網羅的解析方法および新規関節リウマチバイオマーカー

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09700191

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2009548938

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 09700191

Country of ref document: EP

Kind code of ref document: A1