POLYPEPTIDE MARKERS FOR THE DIAGNOSIS OF BLADDER CANCER

摘要

A method for the diagnosis of bladder cancer (BC) and/or for determining a tumor stage of bladder cancer, comprising the step of determining the presence or absence or amplitude of at least six polypeptide markers in a sample, wherein said polypeptide markers are selected from markers 1 to 836, which are characterized by the values for the molecular masses and migration times (CE time).

主权项

1. A method for diagnosing the probability of bladder cancer (BC) in a subject patient comprising:
obtaining a mid-stream urine sample from a subject patient; purifying said urine sample to remove high concentration proteins; separating said urine sample into a plurality of polypeptides; identifying said plurality of polypeptides based on the separation characteristics of each of said plurality of polypeptides in the separating step; comparing said plurality of polypeptides to known polypeptide markers taken from control subjects having BC and not having BC to obtain a subset of polypeptide markers from said plurality of polypeptides that substantially match said known polypeptide markers, wherein said known polypeptide markers are characterized by the following molecular masses and migration times (CE time):NumberMassCE Time14933.9717.224712.2514.331071.5416.343273.4217.751130.3733.361180.5333.77980.3634.0811721.3613.492077.1216.810840.4219.3112115.0422.8123385.4321.1134053.1813.6144240.1314.2151387.049.5162597.4718.6172920.2115.9184509.0925.1198763.4512.1201535.7226.321911.2831.9221191.5434.5233759.9012.8242973.4320.0253325.6715.42610042.8112.2271380.6719.3283502.7012.5293632.6315.9308559.2213.2314960.8615.632858.4118.933847.4020.034980.5118.7351128.5222.4361407.6835.6372093.9231.0382421.0432.5391250.6614.8403242.2018.0411040.5021.2423176.4011.9432778.5532.5441422.5835.8453524.4228.9463831.7424.8472488.7932.2481099.5124.9491936.8229.2501265.6123.4513081.6126.4523266.4826.5533821.9520.4542407.1024.0551596.7519.05610341.7518.1574306.6628.958900.4319.8591645.7415.9601844.5831.4612130.9729.6622359.3531.0633343.5828.5643501.6228.4655675.3119.0661312.6417.6671867.6830.4682186.8531.9692414.5133.6702739.2424.8712907.3633.9723031.4234.0734345.8930.6746211.6714.6751080.5222.9761142.8010.9771321.7410.6781750.7719.5791989.9029.3802946.3832.8814942.3421.4821668.7617.5831688.7114.5841731.7917.7851822.7427.4863870.8330.4874275.4820.6881025.4821.7891353.6722.1901466.6725.2911588.7326.7921878.7227.6932933.3124.1943338.4519.3956055.6315.6962266.0017.2972939.0531.1983021.3918.9991299.6017.61001405.6814.31011526.7219.31021630.7815.41031865.8330.01042687.2425.41052756.2732.81061200.8410.71071391.8110.71081487.6826.11091919.8723.31102063.4814.11112753.4734.21122864.1628.51133858.6521.91144015.9624.41151324.6016.31161793.6015.41172215.7930.71181934.8214.21191935.9613.61202062.8122.912113372.3225.91221495.6818.91231513.4935.41241900.9028.61253108.9433.81261860.8716.21272194.6814.31283956.8121.31291496.7026.51303659.1721.313113169.2425.61321608.7527.51331707.7311.11341592.7217.31352029.8015.013613007.1825.11372117.7729.91382168.8731.01391916.8631.31402802.7834.71411111.7710.81421806.8418.51431594.7126.11441778.7627.01452582.9519.01463605.6321.221473765.3142.771484067.6331.2214910640.419.681781950.8733.91791649.7617.91801312.5526.31811438.4735.21823248.5126.81832742.2625.3184981.6020.71851679.9719.3186801.4322.7187816.4121.05188818.4821.95189838.3625.12190838.4435.05191840.4719.68192852.4720.42193868.4523.35194871.4821.11195875.5121.83196876.4335.13197883.4523.27198884.3624.91199892.3235.22200906.3434.93201906.5121.89202912.5520.04203914.4923.75204915.2835.24205931.5119.99206935.4923.76207936.4921.43208939.5123.792371073.3635.392381078.6919.672391082.5524.372401083.5621.652411096.4135.912421096.5326.122431098.5621.422441099.5621.562451100.5921.622461101.5422.212471106.5226.022481109.6821.062491114.5425.522501114.5721.272511124.5821.042521125.5524.92531125.5821.772541126.5221.22551126.5725.612561134.6323.682571135.5327.512581136.5920.272591137.6524.082601138.6419.552611139.5421.012621140.5321.142631141.5725.22641142.6121.82651143.5636.952661144.4335.572671144.626.25 wherein said CE times are based on capillary electrophoresis using a glass capillary of 90 cm in length with an inner diameter (ID) of 50 μm at an applied voltage of 25 kV, wherein 20% acetonitrile, 0.25% formic acid in water is used as the mobile solvent for the capillary electrophoresis; and wherein said CE times are calibrated relative to the following values:Protein/PolypeptideMigration time (minutes)Aprotinin 9.2Ribonuclease10.9Lysozyme 8.9″REV″ SEQ ID NO: 115.6″ELM″ SEQ ID NO: 223.4″KINCON″ SEQ ID NO: 320.0″GIVLY″ SEQ ID NO: 436.8 determining if said subset of polypeptide markers comprises at least:
a first biomarker selected from the group consisting of: (1) markers 1 to 149 (frequency markers) having (a) a frequency of presence in patients with BC of at least 0.5 and (b) a frequency of presence in patients with BC at least three (3) times the frequency of presence in patients without BC; and (2) markers 1 to 149 (frequency markers) having (a) a frequency of presence in patients without BC of at least 0.5 and (b) a frequency of presence in patients without BC at least three (3) times the frequency of presence in patients with BC,a second biomarker which is different than said first biomarker and is at least one selected from the group consisting of: (1) markers 1 to 149 (frequency markers) having (a) a frequency of presence in patients with BC of at least 0.5 and (b) a frequency of presence in patients with BC at least three (3) times the frequency of presence in patients without BC; and (2) markers 1 to 149 (frequency markers) having (a) a frequency of presence in patients without BC of at least 0.5 and (b) a frequency of presence in patients without BC at least three (3) times the frequency of presence in patients with BC, anda third biomarker which is different than said first and second biomarkers and is at least one selected from the group consisting of markers 1 to 149 (frequency markers); comparing the frequency of presence of said at least said first, second and third biomarkers in said urine sample of said subject patient to the frequency of presence of the same first, second and third biomarkers from said known polypeptide markers taken from said control subjects, ranking said subject patient between said control subjects with BC and without BC based on the second comparing step; and diagnosing the probability of BC in said subject patient based on said ranking.