EP2649450A1 - Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile - Google Patents

Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile

Info

Publication number
EP2649450A1
EP2649450A1 EP11794735.8A EP11794735A EP2649450A1 EP 2649450 A1 EP2649450 A1 EP 2649450A1 EP 11794735 A EP11794735 A EP 11794735A EP 2649450 A1 EP2649450 A1 EP 2649450A1
Authority
EP
European Patent Office
Prior art keywords
markers
sample
polypeptide
bile
polypeptide markers
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.)
Withdrawn
Application number
EP11794735.8A
Other languages
German (de)
English (en)
Inventor
Harald Mischak
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.)
Mosaiques Diagnostics and Therapeutics AG
Original Assignee
Mosaiques Diagnostics and Therapeutics AG
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 Mosaiques Diagnostics and Therapeutics AG filed Critical Mosaiques Diagnostics and Therapeutics AG
Priority to EP11794735.8A priority Critical patent/EP2649450A1/fr
Publication of EP2649450A1 publication Critical patent/EP2649450A1/fr
Withdrawn legal-status Critical Current

Links

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
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
    • H01J49/00—Particle spectrometers or separator tubes
    • H01J49/0027—Methods for using particle spectrometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/08—Hepato-biliairy disorders other than hepatitis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/60—Complex ways of combining multiple protein biomarkers for diagnosis

Definitions

  • the present invention relates to the differentiation of a benign or malignant bile duct stricture from a choledocholithiasis and the differentiation of a cholangiocellular carcinoma from a primary sclerosing cholangitis in the presence of an unclear bile duct stenosis in the bile juice.
  • Bile duct stenoses can have both benign and malignant causes.
  • the occlusion of the bile ducts and the like by stones between donor bile duct and recipient bile duct is rated as benign.
  • the most common stenotic tumors include cholangiocellular carcinoma (CCC), pancreatic carcinoma, hepatocellular carcinoma (HCC) as well as metastases of other carcinomas (Khan et al., J Hepatol 2002, Vol. 37, pages 806-813).
  • Clinical, radiological and endoscopic procedures are used for the diagnosis, but a clear diagnosis without biopsy or surgical procedures is not possible in many cases (Malhi and Gores, Ali ment Pharmacol Ther, 2006, Vol. 23, pages 1287-1296).
  • the CCC is based on the malignant degeneration of cholangiocytes in the entire biliary system and is usually recognized only at an advanced stage, so that less than 50% of patients can be operated on and treated curatively (Malhi and Gores, Aliment Pharmacol Ther, 2006, Vol 23, pages 1287-1296, Singh and Patel, Curr Opin Gastroenterol, 2006, Vol. 22, pages 294-299). Effective chemotherapy does not exist so far. It has already been possible to characterize a risk collective for the CCC.
  • PSC primary sclerosing cholangitis
  • the antineutrophil cytoplasmic antibodies can be found at approx. 85% of patients, but the marker has no high specificity for the PSC and is also found in other diseases, such as in patients with inflammatory bowel disease without PSC.
  • the diagnosis is therefore further based on the representation of the bile ducts with the typical picture of multiple bile duct stenoses.
  • the risk of developing CCC in patients with PSC is greatly increased compared to the normal population: in a large multi-center study from Sweden with 305 PSC patients, a CCC was observed in 8% and 44% of the patients were symptomatic at diagnosis (Broome et al., Gut, 1996, Vol. 38, pages 610-615).
  • ERCP diagnostic or therapeutic ERCP as well as radiological controls (magnetic resonance cholangiopancreatography, MRCP) are performed at regular intervals for surveillance, but to date, all investigations are not sensitive enough for early detection of the CCC.
  • the determination of tumor markers is becoming more important, but these are currently insufficient especially for the early detection of CCC and serve more as a history parameter.
  • the CA19-9 serum marker is currently the most widely used for the diagnosis of CCC, but other tumor or biomarkers are needed for better sensitivity and specificity (Levy et al., Dig Dis Sei, 2005, Vol. 50, Seitel 734-1740, Lempinen et al., J Hepatol, 2007, Vol.47, pages 677-683).
  • the object is achieved by a method for the diagnosis of a benign or malignant bile duct stricture and a CCC comprising the step of determining the presence or absence or amplitude of at least three polypeptide markers in a bile juice sample, wherein the polypeptide marker for the diagnosis of bile duct stricture or of a CCC are selected from the markers which are characterized in Table 1, 2a and Table 2b by values for the molecular mass and the migration time. Table 1. List of markers which, in a multi-marker model, differentiate a benign or malignant bile duct stricture from one
  • Table 2a List of markers which, in a multimarker model, make it possible to differentiate a cholangiocellular carcinoma from benign strictures, in particular sclerosing cholangitis, in body fluid samples, for example in urine
  • the markers are selected from the following markers of Table 2a: Nos. 8503, 13746, 15776, 15800, 16854, 18939, 19773, 20334, 23628, 24393, 25866, 26431, 28103, 28306, 29906, 31480, 32470, 33727, 33840, 33973, 36156, 37056, 37949, 40091, 41514, 42304, 42404, 42833, 45980, 46338, 46649, 48093, 48580, 49958, 50212, 50638, 50904, 51804, 52189, 54687, 58880, 59928, 60259, 61221, 61332, 61984, 64899, 64905, 65746, 69769, 69979, 70413, 72161, 72317, 73434, 73697, 74420, 75025, 76839, 78111, 79720, 81263, 84302, 86426, 87411, 87692
  • Table 2b List of markers which, in a multimarker model, enable the differentiation of a primary sclerosing cholangitis (PSC) from a cholangiocellular carcinoma (CCC), in particular in the case of unclear bile duct steroids in a bile juice sample.
  • PSC primary sclerosing cholangitis
  • CCC cholangiocellular carcinoma
  • the markers are selected from the following markers of Table 2b:
  • amino acid sequence is known. This is listed in Table 3 together with the associated precursor protein.
  • Table 4 Reference values for evaluating the measured presence or absence of markers.
  • ProteinJD mass [Da] CE time [min] Frequency mean log amp (log median)
  • the evaluation of the measured polypeptides can be based on the presence or absence or amplitude of the markers taking into account the following limits:
  • Specificity is defined as the number of actual negative samples divided by the sum of the number of actual negatives and the number of false positives. A specificity of 100% means that a test identifies all healthy persons as healthy, i. no healthy person is identified as ill. This does not say anything about how well the test detects sick patients.
  • Sensitivity is defined as the number of actually positive samples divided by the sum of the number of actual positives and the number of false negatives. A sensitivity of 100% means that the test detects all patients. He does not say how well the test detects healthy people.
  • the markers according to the invention make it possible to achieve a specificity of at least 65%, preferably at least 75%, more preferably 85%, for the recognition of a biliary stricture or of a CCC.
  • the markers according to the invention make it possible to achieve a sensitivity of at least 65%, preferably at least 75%, more preferably 85%, for the recognition of a biliary stricture or of a CCC.
  • the migration time is determined by capillary electrophoresis (CE) - such. B. in example under point 2 - determined.
  • CE capillary electrophoresis
  • the eluent used is, for example, 20% acetonitrile, 0.25% formic acid in water.
  • CE migration time can vary. Nevertheless, the order in which the polypeptide markers elute is typically the same for each CE system used under the conditions indicated. To even out any differences in migration time, the system can be normalized using standards for which migration times are known. These standards may, for. Example, the polypeptides given in the examples be (see example point 3). Variation in CE time is relatively small between individual measurements, typically in the range of ⁇ 2 min, preferably in the range of ⁇ 1 min, more preferably ⁇ 0.5 min, even more preferably ⁇ 0.2 min or 0.1 minute
  • the characterization of the polypeptides shown in Tables 1 to 5 was determined by capillary electrophoresis mass spectrometry (CE-MS), a method which was described e.g. In detail by Neuhoff et al. ⁇ Rapid Communications in mass spectrometry, 2004, Vol. 20, pages 149-156).
  • CE-MS capillary electrophoresis mass spectrometry
  • the variation of molecular masses between individual measurements or between different mass spectrometers is relatively small with exact calibration, typically in the range of ⁇ 0.1%, preferably in the range of ⁇ 0.05%, more preferably ⁇ 0.03%, even more preferably ⁇ 0.01% or 0.005%.
  • polypeptide markers according to the invention are proteins or peptides or degradation products of proteins or peptides. They can be chemically modified, for. B. by post-translational modifications such as glycolization, phosphorylation, alkylation or disulfide bridging, or by other reactions, e.g. B. in the context of degradation, be changed.
  • polypeptide markers molecular mass and migration time
  • polypeptides of the invention are used to differentiate a benign or malignant bile duct stricture from choledocholithiasis and to differentiate a CCC from a PSC in unclear bile duct stenosis.
  • Diagnosis is the process of gaining knowledge by assigning symptoms or phenomena to a disease or injury.
  • the presence or absence of certain polypeptide markers is also used for diagnosis.
  • the presence or absence of a polypeptide marker can be measured by any method known in the art. Methods that can be used are exemplified below.
  • a polypeptide marker is present when its reading is at least as high as the threshold. If its reading is below that, the polypeptide marker is absent.
  • the threshold value can either be determined by the sensitivity of the measurement method (detection limit) or defined based on experience.
  • the threshold is preferably exceeded when the sample reading for a given molecular mass is at least twice that of a blank (eg, only buffer or solvent).
  • the polypeptide marker (s) is / are used to measure its presence or absence, the presence or absence being indicative of the diagnosis of benign or malignant bile duct stricture or CCC.
  • polypeptide markers which are typically present in individuals with a benign or malignant bile duct stricture or in individuals with a CCC, but in individuals without a bile duct stricture, e.g. As those with Choledocholithiasis, or a CCC, z.
  • those with PSC rarely or not occur.
  • polypeptide markers that are present in patients with bile duct stenosis of a different etiology but are not or only rarely present in patients with PSC or CCC.
  • amplitude markers can also be used for diagnosis.
  • Amplitude markers are used in such a way that it is not the presence or absence that is decisive, but the height of the signal (the amplitude) decides in the presence of the signal in both groups.
  • the tables show the average equivalences of the corresponding signals (characterized by mass and migration time) over all measured samples. Two standardization methods are possible in order to achieve comparability between differently concentrated samples or different measurement methods. In the first approach, all peptide signals of a sample are sampled to a total amplitude of 1 million Counts standardized. The respective mean amplitudes of the single markers are therefore given as parts per million (ppm).
  • amplitude markers via an alternative standardization procedure: in this case, all peptide signals of a sample are scaled with a common normalization factor. For this purpose, a linear regression is formed between the peptide amplitudes of the individual samples and the reference values of all known polypeptides. The slope of the regression line just corresponds to the relative concentration and is used as a normalization factor for this sample.
  • the decision to make a diagnosis depends on how high the amplitude of the respective polypeptide markers in the patient sample is in comparison to the mean amplitudes in the reference groups or the "sick" group. If the value is close to the mean amplitude of the "sick" group, the polypeptide markers for the detection of a ganglion stricture are characterized by the presence of a benign or malignant stricture and the absence of choledocholithiasis and in the polypeptide markers for the recognition of a CCC by the CCC presence in the presence of a PSC (CCC on top of PSC), but also the presence of a CCC in the absence of a PSC.
  • the presence of a benign or malignant stricture in the polypeptide markers for the detection of a benign or malignant stricture stricture and CCC in the case of the polypeptide markers is not to be assumed.
  • the distance to the mean amplitude can be interpreted as a probability of belonging to a group.
  • the distance between the measured value and the mean amplitude may be considered as a probability of belonging to a group.
  • a frequency marker is a variant of the amplitude marker, in which the amplitude is low in some samples. It is possible to convert such frequency markers into amplitude markers in which, in the calculation of the amplitude, the corresponding samples in which the marker is not found, with a very small amplitude - in the range of the detection limit - are included in the calculation.
  • the individual from whom the sample is derived, in which the presence or absence of one or more polypeptide markers is determined may be any individual in whom a benign or malignant genetic disorder or CCC may occur.
  • the subject is a mammal, most preferably a human.
  • the sample measuring the presence or absence of the polypeptide marker (s) of the invention may be any sample consisting of the body of the individual is won.
  • the sample is a sample having a polypeptide composition suitable for making statements about the condition of the individual.
  • it may be blood, urine, synovial fluid, tissue fluid, body secretions, sweat, cerebrospinal fluid, lymph, intestinal, gastric, pancreatic, bile, tears, tissue, sperm, vaginal fluid, or a stool sample.
  • it is a liquid sample.
  • the sample is a bile juice sample.
  • the markers of Table 2b are suitable for diagnosis. If other samples are used, the markers of Table 2a are suitable. These markers are particularly suitable when a urine sample is used as a sample. Urine samples are easier to obtain compared to bile juice samples. However, bile juice samples seem to have a better informative value.
  • the urine samples are first diagnosed. If the results are unclear, then further analyzes based on bile juice samples are carried out.
  • Bile juice samples may be known as known in the art.
  • a bile juice extraction is used in the course of an endoscopic procedure.
  • the bile juice sample may be e.g. be removed from the bile duct by means of an endoscopically inserted catheter or by means of another apparatus.
  • the presence or absence of a polypeptide marker in the sample can be determined by any method known in the art suitable for measuring polypeptide markers. Those skilled in such methods are known. In principle, the presence or absence of a polypeptide marker can be determined by direct methods such as e.g. Mass spectrometry, or indirect methods, e.g. by ligands.
  • the sample of the subject eg the bile juice sample
  • the treatment may include, for example, purification, separation, dilution or concentration.
  • the methods may be, for example, a centrifugation, filtration, ultrafiltration, dialysis, precipitation or chromatographic methods such as affinity separation or separation by means of ion exchange chromatography, or an electrophoretic separation.
  • the sample is separated by electrophoresis prior to its measurement, purified by ultracentrifugation and / or separated by ultrafiltration into fractions containing polypeptide markers of a certain molecular size.
  • a mass spectrometric method is used to determine the presence or absence of a polypeptide marker, which method may precede purification or separation of the sample.
  • the mass spectrometric analysis has the advantage over current methods that the concentration of many (> 100) polypeptides of a sample can be determined by a single analysis. Any type of mass spectrometer can be used. With mass spectrometry, it is possible to routinely 10 fmol of a polypeptide marker, ie 0.1 ng of a 10 kDa protein with a measurement accuracy of approx. ⁇ 0.01% from a complex mixture. In mass spectrometers, an ion-forming unit is coupled to a suitable analyzer.
  • electrospray ionization (ESI) interfaces are most commonly used to measure ions from liquid samples, whereas the matrix assisted laser desorption / ionization (MALDI) technique is used to measure ions from a sample crystallized with a matrix , For analysis of the resulting ions z.
  • MALDI matrix assisted laser desorption / ionization
  • quadrupoles ion traps or time-of-flight (TOF) analyzers can be used.
  • electrospray ionization the molecules present in solution are inter alia. sprayed under the influence of high voltage (eg 1-8 kV) to form charged droplets, which become smaller due to evaporation of the solvent. Finally, so-called Coulomb explosions lead to the formation of free ions, which can then be analyzed and detected.
  • high voltage eg 1-8 kV
  • TOF analyzers have a very high scanning speed and achieve a very high resolution.
  • Preferred methods for determining the presence or absence of polypeptide starch include gas phase ionization spectroscopy, such as laser absorption / ionization mass spectrometry, MALDI-TOF-MS, SELDI-TOF-MS ( Surface-enhanced laser desorption ionization), LC-MS (liquid chromatographic mass spectrometry), 2D-PAGE-MS and capillary electrophoresis mass spectrometry (CE-MS). All of the methods mentioned are known to the person skilled in the art.
  • gas phase ionization spectroscopy such as laser absorption / ionization mass spectrometry, MALDI-TOF-MS, SELDI-TOF-MS ( Surface-enhanced laser desorption ionization), LC-MS (liquid chromatographic mass spectrometry), 2D-PAGE-MS and capillary electrophoresis mass spectrometry (CE-MS). All of the methods mentioned are known to the person skilled in the art.
  • CE-MS in which the capillary electrophoresis is coupled with mass spectrometry. This method is in detail z.
  • the CE-MS technique allows the presence of a few hundred polypeptide markers of a sample simultaneously in a short time, with a low volume and high sensitivity. vote. After a sample has been measured, a pattern of the measured polypeptide markers is prepared.
  • CE-MS method in which the CE is coupled online to an ESI-TOF-MS.
  • solvents include acetonitrile, methanol and the like.
  • the solvents may be diluted with water and an acid (eg, 0.1% to 1% formic acid) added to protonate the analyte, preferably the polypeptides.
  • Capillary electrophoresis makes it possible to separate molecules according to their charge and size. Neutral particles migrate at the rate of electroosmotic flow upon application of a current, cations are accelerated to the cathode and anions are retarded.
  • the advantage of capillaries in electrophoresis is the favorable ratio of surface to volume, which allows a good removal of the Joule heat arising during the current flow. This in turn allows the application of high voltages (usually up to 30 kV) and thus a high separation efficiency and short analysis times.
  • quartz glass capillaries are used with internal diameters of typically 50 to 75 ⁇ . The used lengths are 30-100 cm.
  • the capillaries are usually made of plastic around coated quartz glass.
  • the capillaries may be both untreated, i. on the inside show their hydrophilic groups, as well as be coated on the inside. A hydrophobic coating can be used to improve the resolution.
  • a pressure which is typically in the range of 0-1 psi may also be applied. The pressure can also be created during the separation or changed during the process.
  • the markers of the sample are separated by capillary electrophoresis, then directly ionized and transferred online to a mass spectrometer coupled thereto for detection.
  • polypeptide markers can advantageously be used for diagnostics.
  • Preferred is the use of at least 5, 6, 8, or 10 markers.
  • 20 to 50 markers are used.
  • Bile juice was used to detect polypeptide markers for diagnosis. Bile fluid was taken from patients with choledocholithiasis, patients with PSC, patients with CCC and patients with CCC on top of PSC.
  • the lipids present in high concentration in the bile juice were precipitated by addition of 1-butanol and diisopropyl ether and all the macromolecular bile juice components (> 10 kDa) were separated by ultrafiltration.
  • 700 .mu.l bile juice were removed and pipetted to 700 .mu.l of a 1-butanol / diisopropyl ether (4: 6, v / v) mixture.
  • the sample was then mixed on the vortex shaker until a homogeneous yellowish emulsion formed. After centrifugation at 13,000 rpm and 4 ° C.
  • the aqueous phase from the delipidation was mixed with 500 ⁇ 8M (w / v) urea solution and added to the UF filter (10 kDa MWCO, Sartorius, Göttingen, D). Then 1.0 ml of dist. H 2 0 was added and the UF was carried out at 3000 rpm in a centrifuge until 1.1 ml of ultrafiltrate were obtained. The resulting 1.1 ml of filtrate was then applied to a PD 10 column (GE Healthcare, Munich, D) and eluted with 2.5 ml of 0.01% NH 4 OH and lyophilized. For CE-MS measurement, the polypeptides were then resuspended in 20 ⁇ H 2 O (HPLC grade, Merck).
  • CE-MS measurements were performed with a capillary electrophoresis system from Beckman Coulter (P / ACE MDQ System, Beckman Coulter Inc, Fullerton, USA) and a Bruker ESI-TOF mass spectrometer (micro-TOF MS, Bruker Daltonik, Bremen, D).
  • the CE capillaries were purchased from Beckman Coulter, they had an ID / OD of 50/360 ⁇ and a length of 90 cm.
  • the mobile phase for the CE separation consisted of 20% acetonitrile and 0.25% formic acid in water.
  • the coupling of CE and MS was realized by a CE-ESI-MS Sprayer Kit (Agilent Technologies, Waldbronn, DE).
  • the duration of the injection was 99 seconds. With these parameters about 150 nl of the sample were injected into the capillary, this corresponds to about 10% of the capillary volume.
  • a "stacking" technique was used, injecting an IMNH 3 solution for 7 sec (at 1 psi) before injecting the sample and injecting a 2M formic acid solution for 5 sec after sample injection Separation voltage (30 kV), the analytes are automatically concentrated between these solutions.
  • the following CE separation was performed with a pressure method: 0 psi for 40 minutes, 0.1 psi for 2 minutes, 0.2 psi for 2 minutes, 0.3 psi for 2 minutes, 0.4 psi for 2 minutes, finally 32 min at 0.5 psi.
  • the total duration of a separation run was thus 80 minutes.
  • the "Nebulizer gas” was set to the lowest possible value.
  • the voltage applied to the spray needle to generate the electrospray was 3700 - 4100 V.
  • the other settings on the mass spectrometer were optimized according to the manufacturer's instructions for a peptide detection. The spectra were recorded over a mass range of m / z 400 to m / z 3000 and accumulated every 3 seconds.
  • the proteins / polypeptides are each used in a concentration of 10 pmol / ⁇ in water.
  • the polypeptides designated with their amino acid sequence are synthetic peptides.
  • the molecular masses of the protein / polypeptide standards are given in Table 4.
  • the most probable assignment is that in which there is a substantially linear relationship between the shift for the peptide 1 and for the peptide 2.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Biophysics (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne un procédé de diagnostic d'un rétrécissement bénin ou malin des voies biliaires et/ou d'un cholangiocarcinome (CCC), comprenant l'étape consistant à détecter au moins trois marqueurs polypeptidiques dans un liquide corporel, le marqueur polypeptidique faisant partie des marqueurs caractérisés dans le tableau 1 et/ou le tableau 2a et/ou 2b par des valeurs de masse molaire et de temps de migration.
EP11794735.8A 2010-12-10 2011-12-12 Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile Withdrawn EP2649450A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP11794735.8A EP2649450A1 (fr) 2010-12-10 2011-12-12 Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP10194613 2010-12-10
EP11174088 2011-07-15
EP11794735.8A EP2649450A1 (fr) 2010-12-10 2011-12-12 Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile
PCT/EP2011/072441 WO2012076723A1 (fr) 2010-12-10 2011-12-12 Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile

Publications (1)

Publication Number Publication Date
EP2649450A1 true EP2649450A1 (fr) 2013-10-16

Family

ID=45346474

Family Applications (1)

Application Number Title Priority Date Filing Date
EP11794735.8A Withdrawn EP2649450A1 (fr) 2010-12-10 2011-12-12 Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile

Country Status (3)

Country Link
US (1) US20140027283A1 (fr)
EP (1) EP2649450A1 (fr)
WO (1) WO2012076723A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3042203B1 (fr) * 2013-09-06 2017-11-15 Leibniz - Institut für Analytische Wissenschaften - ISAS - E.V. Biomarqueurs du carcinome cholangiocellulaire (ccc)

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10021737C2 (de) 2000-05-04 2002-10-17 Hermann Haller Verfahren und Vorrichtung zur qualitativen und/oder quantitativen Bestimmung eines Protein- und/oder Peptidmusters einer Flüssigkeitsprobe, die dem menschlichen oder tierischen Körper entnommen wird
AU2003253329A1 (en) * 2002-07-25 2004-02-16 Sabine Debuschewitz Molecular markers for cholangiolar carcinoma
WO2006042186A2 (fr) * 2004-10-07 2006-04-20 Emory University Preparation pour examen gastro-intestinal
WO2010031822A1 (fr) * 2008-09-17 2010-03-25 Mosaiques Diagnostics And Therapeutics Ag Carcinome à cellules rénales

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2012076723A1 *

Also Published As

Publication number Publication date
WO2012076723A1 (fr) 2012-06-14
US20140027283A1 (en) 2014-01-30

Similar Documents

Publication Publication Date Title
EP2338054A1 (fr) Carcinome à cellules rénales
DE102011017084B4 (de) Massenspektrometriedaten-Erfassungsmodus zur Erzielung einer zuverlässigeren Proteinquantifizierung
EP2255203A2 (fr) Procédé et marqueur permettant de diagnostiquer des maladies et dommages tubulaires rénaux
EP1917531A2 (fr) Marqueurs polypeptidiques pour le diagnostic du cancer de la vessie
WO2008110593A2 (fr) Procédé et marqueurs pour le diagnostic de maladies rénales
EP1955064A1 (fr) Marqueur polypeptidique pour diagnostiquer et evaluer la stenose du meat de l'uretre
EP2437064B1 (fr) Biomarqueur d'hépatite
WO2011000938A1 (fr) Procédé et marqueur permettant de diagnostiquer une insuffisance rénale aiguë
EP2317320A2 (fr) Marqueurs polypeptidiques pour la diagnose du cancer de la prostate
CN111936851A (zh) 用于检测维生素d代谢物的方法和系统
EP2478377A2 (fr) Polypeptide marqueur pour le diagnostic et l'évaluation de maladies vasculaires
EP2212703A1 (fr) Procédé et marqueurs pour diagnostiquer le diabète sucré
WO2013113744A2 (fr) Marqueurs polypeptidiques pour le diagnostic et l'évaluation de l'insuffisance cardiaque
WO2009047280A2 (fr) Marqueur polypeptidique pour le diagnostic du cancer de la prostate
EP2394171A2 (fr) Maladie polykystique rénale autosomique dominante (adpkd)
EP2649450A1 (fr) Procédé et marqueur pour le diagnostic d'un rétrécissement des voies biliaires et d'un cholangiocarcinome à partir de la bile
EP1896856A1 (fr) Marqueur polypeptidique servant a detecter le rejet d'un rein greffe a un stade precoce
WO2012123527A1 (fr) Procédé et marqueurs de diagnostic de formes infracliniques et cliniques du rejet tubulo-interstitiel à médiation lymphocytaire t après une transplantation rénale
EP1784649B1 (fr) Marqueurs polypeptidiques pour le diagnostic de l'arteriosclerose
WO2006106115A2 (fr) Marqueur de polypeptide pour le diagnostic d'alzheimer
WO2013150132A2 (fr) Marqueurs polypeptidiques pour diagnostiquer et évaluer des accidents vasculaires cérébraux
WO2007063089A2 (fr) Marqueur polypeptidique pour diagnostiquer et evaluer des maladies vasculaires

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20130705

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAX Request for extension of the european patent (deleted)
17Q First examination report despatched

Effective date: 20150506

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20160701