WO2016195051A1 - Panel de biomarqueurs de plasma permettant le diagnostic du cancer du pancréas - Google Patents

Panel de biomarqueurs de plasma permettant le diagnostic du cancer du pancréas Download PDF

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WO2016195051A1
WO2016195051A1 PCT/JP2016/066514 JP2016066514W WO2016195051A1 WO 2016195051 A1 WO2016195051 A1 WO 2016195051A1 JP 2016066514 W JP2016066514 W JP 2016066514W WO 2016195051 A1 WO2016195051 A1 WO 2016195051A1
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biomarker
pancreatic cancer
present
antibody
cancer
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Japanese (ja)
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一文 本田
哲司 山田
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国立研究開発法人国立がん研究センター
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    • 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
    • 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
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N37/00—Details not covered by any other group of this subclass

Definitions

  • the present invention relates to a plasma biomarker panel for diagnosing pancreatic cancer and a method for detecting pancreatic cancer using the plasma biomarker.
  • Pancreatic cancer is an intractable cancer, and the 5-year survival rate is very low compared to other solid cancers. This is because early pancreatic cancer is difficult to detect and is often detected as advanced cancer, so there are few cases where radical surgery can be performed. The development of blood diagnostic markers that efficiently detect early pancreatic cancer that can be radically operated is likely to increase the early pancreatic cancer detection rate and improve pancreatic cancer mortality.
  • CA19-9 does not respond to patients with early pancreatic cancer.
  • Lewis antigen-negative patients do not produce CA19-9, an increase in CA19-9 is not detected even if Lewis antigen-negative patients are advanced pancreatic cancer patients.
  • ASCO the significance of CA19-9 as a diagnostic biomarker for early pancreatic cancer has been denied.
  • Non-patent Document 1 Nakamura et.al, table 1
  • mRNA is extracted from cells obtained by microdigesting a normal pancreatic duct and an invasive pancreatic duct cancer portion derived from a surgical specimen, and hybridized to a genome wide cDNA microarray to find the mRNA. Since these mRNAs were found based on changes in expression in tissues in which cancer occurs, it is necessary to use tissues collected from subjects in order to use them as diagnostic biomarkers for pancreatic cancer There is.
  • An object of the present invention is to provide a method for detecting pancreatic cancer using a body fluid. It is another object of the present invention to provide a biomarker capable of non-invasively detecting pancreatic cancer.
  • pancreatic cancer patient plasma and healthy subject plasma were analyzed as a Receiver Operating Characteristic.
  • ROC analysis aria of under curve (AUC)
  • AUC aria of under curve
  • 23 antibodies exceeding 0.8 were found. This means that the binding amount of each antibody has a highly statistically significant difference between pancreatic cancer patients and healthy individuals. That is, these 23 antibodies and the 23 proteins recognized by the respective antibodies or fragments thereof were found to be biomarkers capable of noninvasively detecting pancreatic cancer, thereby completing the present invention.
  • the present invention relates to the following.
  • detecting and / or quantifying at least one biomarker in a body fluid sample from the subject The biomarkers are AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6, ATP6AP1, and A method for detecting pancreatic cancer, which is a protein selected from the group consisting of HYOU1 or a fragment thereof.
  • the method according to [1] wherein the body fluid sample is whole blood, serum, plasma, or urine.
  • the immunological assay includes Western blot, dot blot analysis, slot blot analysis, radioimmunoassay (RIA), peptide microarray, enzyme immunoassay (EIA), and enzyme-linked immunosorbent assay ( The method according to [3], which is selected from the group consisting of (ELISA).
  • ELISA enzyme immunoassay
  • [6] The method according to [5], comprising a step of bringing the body fluid sample into contact with the antibody, and a step of detecting and / or quantifying the binding between the antibody and at least one biomarker in the body fluid sample. .
  • [7] The method according to [1] or [2], wherein the biomarker is detected and / or quantified using a mass spectrometer.
  • [8] The method according to any one of [1] to [7] for use in diagnosis of pancreatic cancer.
  • [9] The method according to any one of [1] to [8], wherein the subject has pancreatic cancer.
  • the method further comprises the step of comparing the level of the biomarker in the body fluid sample with the control level of the biomarker; (I) AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6 in the subject sample , When at least one biomarker level selected from ATP6AP1 is higher than the control level, and / or (ii) when the HYOU1 level in the subject sample is lower than the control level, The method according to any one of [1] to [8], wherein pancreatic cancer is used as an indicator.
  • a diagnostic agent for pancreatic cancer containing an antibody against a biomarker The biomarkers are AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6, ATP6AP1, and A diagnostic agent for pancreatic cancer, which is at least one protein selected from the group consisting of HYOU1 or a fragment thereof.
  • the diagnostic agent according to [11] wherein the body fluid sample is whole blood, serum, plasma or urine.
  • a pancreatic cancer detection kit containing an antibody against a biomarker The biomarkers are AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6, ATP6AP1, and A kit which is at least one protein selected from the group consisting of HYOU1 or a fragment thereof.
  • a biomarker for detecting pancreatic cancer comprising at least one protein selected from the group consisting of the fragments or a fragment thereof.
  • the present invention provides a biomarker for detecting pancreatic cancer, a method for detecting pancreatic cancer using the biomarker, a kit for detecting pancreatic cancer or a diagnostic agent containing an antibody against the biomarker.
  • the biomarker of the present invention can be used for efficiently discriminating pancreatic cancer, and a method for detecting or diagnosing pancreatic cancer using these biomarkers is extremely effective clinically.
  • the present invention detects pancreatic cancer by detecting and / or quantifying a biomarker using a body fluid sample derived from a subject.
  • Such a non-invasive biomarker invention provides a new medical strategy for screening for pancreatic cancer, and thus has the potential to contribute to a reduction in pancreatic cancer mortality.
  • the biomarker can be detected by an immunological assay method such as ELISA, or a clinical test method using mass spectrometry or the like.
  • a biomarker panel for diagnosing pancreatic cancer can be provided by combining the biomarker of the present invention or by combining with a known biomarker.
  • FIG. 1 shows the results of hybridizing an anti-AP3B1 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 2 shows the results of hybridizing an anti-C2 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 3 shows the results of hybridizing an anti-CKS1B antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 4 shows the results of hybridizing an anti-CKS2 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 5 shows the results of hybridizing an anti-CSPG2 antibody to a digestive cancer plasma microarray.
  • FIG. 6 shows the results of hybridizing an anti-CYCS antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 7 shows the results of hybridizing an anti-CD82 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 8 shows the results of hybridizing anti-PI3 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 9 shows the results of hybridizing an anti-RNASE1 antibody to a digestive cancer plasma microarray.
  • FIG. 10 shows the results of hybridizing an anti-RNASET2 antibody to a digestive cancer plasma microarray.
  • FIG. 11 shows the results of hybridizing an anti-VRK2 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 12 shows the results of hybridizing an anti-EV1 antibody to a digestive cancer plasma microarray.
  • FIG. 13 shows the results of hybridizing an anti-HMGB2 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 14 shows the results of hybridizing an anti-MST4 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 15 shows the results of hybridizing an anti-MMP9 antibody to a digestive cancer plasma microarray.
  • FIG. 16 shows the results of hybridizing an anti-MYBL2 antibody to a digestive cancer plasma microarray.
  • FIG. 17 shows the results of hybridizing an anti-HYOU1 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 18 shows the results of hybridizing an anti-PPM1B antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 19 shows the results of hybridizing an anti-AK3L1 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 20 shows the results of hybridizing an anti-IGFBP2 antibody to a digestive cancer plasma microarray.
  • FIG. 21 shows the results of hybridizing anti-STMN1 antibody to a digestive cancer plasma microarray.
  • FIG. 22 shows the results of hybridizing an anti-ANXA6 antibody to a gastrointestinal cancer plasma microarray.
  • FIG. 23 shows the results of hybridizing an anti-ATP6AP1 antibody to a
  • the present invention relates to a method for detecting pancreatic cancer.
  • 23 types of proteins or fragments thereof identified by proteomic analysis using an antibody library and a plasma microarray were subjected to Receiver Operating Characteristic (ROC) analysis for discriminating pancreatic cancer from body fluids of healthy subjects. It has an area under curve (AUC) value greater than 0.8.
  • AUC value of 0.8 or greater means a clinically useful diagnostic marker.
  • these proteins or fragments thereof can be biomarkers for risk of developing, detecting, diagnosing and monitoring pancreatic cancer.
  • the biomarker of the present invention may be used to detect not only pancreatic cancer but also malignant tumors occurring in the pancreas.
  • the biomarker of the present invention may be used for diagnosing precancerous lesions or risk diseases of pancreatic cancer.
  • Pancreatic cancer can be detected using a substance that binds to the biomarker of the present invention.
  • the present invention can detect pancreatic cancer by detecting and / or measuring the presence of a biomarker in the body fluid of a subject.
  • the present invention can also detect and / or measure biomarkers at the protein level by immunological assays or mass spectrometers.
  • the method of the present invention can detect pancreatic cancer non-invasively.
  • early pancreatic cancer can be detected.
  • the biomarker of the present invention may be used alone, or may be used as a panel combined with a known biomarker or a panel combined with the biomarker of the present invention.
  • the biomarker of the present invention is a protein or a fragment thereof, and can be detected by an immunoassay method such as ELISA or a mass spectrometry method, and thus has utility in clinical medicine, in-vitro diagnosis and clinical examination fields. It is expected to be demonstrated.
  • the present invention relates to a method for detecting pancreatic cancer by detecting and / or quantifying an elevated or lowered biomarker in a body fluid sample such as urine, blood (eg, whole blood, serum or plasma).
  • a body fluid sample such as urine, blood (eg, whole blood, serum or plasma).
  • the biomarker of the present invention is a protein or a fragment thereof.
  • the present invention also relates to a kit or diagnostic agent for detecting pancreatic cancer.
  • the kit or diagnostic agent of the present invention can be used to carry out the method of the present invention.
  • the method of the present invention is performed by an immunological assay using an antibody or the like.
  • the method of the invention comprises (a) contacting a body fluid sample from a subject with the antibody, (b) binding of at least one biomarker in the body fluid sample and the antibody. Detecting and / or quantifying pancreatic cancer, and optionally comparing the level of the biomarker in the body fluid sample obtained in (c) (b) with a control level of said biomarker It is a method to do.
  • the difference between the level of the biomarker in the subject and the control level can be an indicator of the presence of pancreatic cancer.
  • Biomarker of the Present Invention a biomarker is associated with cancer, and its detection or measurement is a substance useful in cancer detection, patient diagnosis or monitoring.
  • the biomarker in the present invention is a protein or a fragment thereof.
  • the biomarker of the present invention is to detect cancer, to diagnose pancreatic cancer including early diagnosis, to diagnose precancerous lesions, to diagnose the occurrence of malignant tumors in the pancreas, Screening healthy or high-risk populations for the presence of pancreatic cancer, determining the prognosis of the subject, and the course of the subject while undergoing surgery, radiation, chemotherapy or other cancer treatment It can be used for various purposes such as monitoring.
  • biomarker for detecting pancreatic cancer plasma circulating in the plasma of pancreatic cancer patients was measured using a plasma microarray and an antibody library developed by the present inventors.
  • the identified biomarkers of the present invention are listed in Table 1. All 23 identified proteins have an AUC value greater than 0.8 (Table 2).
  • a gastrointestinal cancer plasma microarray is prepared, the antibodies in the antibody library are hybridized, and the fluorescence is obtained. The strength was measured and digitized. As a result, 23 types of antibodies that discriminate between pancreatic cancer patients and healthy individuals with an AUC value of 0.8 or more were identified.
  • Proteins that show significantly higher levels in the blood of pancreatic cancer patients compared to healthy subjects are AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, They were MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6, ATP6AP1.
  • the protein showing a significantly lower level in the blood of pancreatic cancer patients compared to the healthy subject sample was HYOU1 (FIGS. 1 to 23).
  • biomarker of the present invention refers to at least one of the proteins shown in Table 1 or fragments thereof.
  • the term includes all homologues of biomarkers in humans, naturally occurring allelic variant products, isoforms and precursors. Allelic variants may contain conservative amino acid substitutions from the amino acid sequences of the biomarkers of the invention and include amino acid substitutions derived from corresponding positions in the biomarker homologues of the invention.
  • each transcriptional variant generated by alternative splicing is also included in the biomarker of the present invention.
  • protein fragments are also included in the biomarkers of the present invention.
  • a fragment is, for example, a peptide of 3, 4, 5, 6, 7, 8, 9 or 10 amino acids or more.
  • the biomarkers of the present invention also include proteins or fragments thereof in species other than human that correspond to biomarkers in humans. Since the biomarker of the present invention can be used for detection of pancreatic cancer, it is also referred to as pancreatic cancer marker or cancer biomarker. Moreover, peptide sequences and sugar chain structures recognized by antibodies that bind to the biomarkers of the present invention are also included in the biomarkers of the present invention. Accordingly, a method for detecting pancreatic cancer by detecting or quantifying the peptide sequence and sugar chain is also included in the present invention.
  • pancreatic cancer of the present invention refers to all neoplastic cell growth and proliferation and all precancerous and cancerous cells and tissues.
  • pancreatic cancer is not limited to serous cystadenocarcinoma, mucinous cystadenocarcinoma, intraductal lactose mucinous adenocarcinoma, intraductal tubular adenocarcinoma, (pancreatic) atypical hyperplasia and Intraepithelial carcinoma, invasive pancreatic duct cancer, acinar cell carcinoma, high molecular endocrine cancer, small molecule endocrine cancer (small cell cancer), coexisting tumor (exocrine / endocrine), pancreatic duct cancer and islet cell mixed cancer, pancreatic duct cancer Includes mixed islet and acinar cell carcinoma, solid pseudopapillary tumor, pancreatic blastoma, anaplastic carcinoma, lei
  • Invasive pancreatic liver cancer includes lactose adenocarcinoma, tubular adenocarcinoma, adenosquamous carcinoma, mucinous cancer, anaplastic cancer, invasive mucinous cystadenocarcinoma, and invasive cancer derived from intraductal tumor.
  • Tubular adenocarcinoma includes well differentiated tubular adenocarcinoma, moderately differentiated tubular adenocarcinoma and low molecular tubular adenocarcinoma.
  • the cancer detected using the method, diagnostic agent and kit of the present invention may be a non-primary malignant tumor.
  • Such malignant tumors include, but are not limited to, for example, breast cancer (eg, invasive (invasive), preinvasive, inflammatory, Paget's disease, metastatic or recurrent); gastrointestinal cancer / digestive tract Cancer (eg, appendix, bile duct, colon, esophagus, gallbladder, gastric, intestine, liver, pancreas, kidney and stomach); genitourinary cancer / urinary cancer For example, adrenal gland, bladder, kidney cancer, penis, prostate, testicles and urinary organs; gynecological cancer (eg, cervix, endometrium, fallopian tube, ovary, uterus, vagina and vulva); head and neck cancer (eg, , Eyes, head and neck, lower jaw, pharynx, nasal cavity, oral cancer, pharynx, salivary gland, sinuses, throat, thyroid, tongue and tonsils); hematological cancer / Blood cancer (eg, Hodgkin's disease
  • the cancer is ovarian cancer.
  • the cancer is selected from the group consisting of breast cancer, endometrial cancer, cervical cancer, lung cancer, colon cancer, prostate cancer, melanoma, glioblastoma, sarcoma, bladder cancer and head and neck cancer It may be a cancer.
  • cancer detected using the method, diagnostic agent and kit of the present invention may include precancerous lesions of pancreatic cancer, diabetes, chronic pancreatitis, hereditary pancreatitis, intraductal papillary mucinous tumor, and pancreatic cyst .
  • subject is a warm-blooded animal, eg, a mammal.
  • subject is preferably a human.
  • Body fluids include but are not limited to urine, whole blood, plasma, serum, tears, semen, saliva, sputum, exhaled breath, nasal discharge, pharyngeal exudate, bronchoalveolar lavage, tracheal aspiration, interstitial fluid, lymph fluid, Meningeal fluid, amniotic fluid, glandular fluid, feces, sweat, mucous, vaginal or urethral secretions, cerebrospinal fluid and percutaneous exudate, preferably whole blood, plasma, serum, urine, sweat More preferably, it is plasma.
  • the biomarker of the present invention is preferably detected in vitro in human plasma.
  • the sample used in the method of the present invention may be obtained directly from a source, or may be used after pretreatment by a method known to those skilled in the art.
  • the sample may be pretreated in any manner and / or prepared in any convenient medium that does not interfere with the assay.
  • the sample may be used after sample processing such as filtration, distillation, extraction, concentration, inactivation of interference components, addition of reagents, and the like.
  • the biomarker of the present invention may be contained in exosomes. Therefore, in the present invention, exosomes isolated from body fluids or exosomes in body fluids may be used as samples.
  • the biomarker of the present invention may be bound or contained in a tumor cell (CTC) circulating in a body fluid. Therefore, in the present invention, CTC in body fluid or CTC isolated from body fluid can be used as a sample. Moreover, pancreatic cancer can be detected by detecting proteins and metabolites from CTC and detecting DNA fragments circulating in the blood.
  • CTC tumor cell
  • the term “detect” or “detect” includes assaying for the presence or absence of a target biomarker protein or fragment thereof of the present invention, establishing an assay system, or determining.
  • the term “detect” or “detect” includes quantitative, semi-quantitative and qualitative detection methods.
  • the detection method is preferably an immunological method such as ELISA or a method using a mass spectrometer.
  • the detection method provides an output (ie, readout or signal) that includes information regarding the presence, absence or amount of the biomarker of the invention in a sample obtained from a subject. Is preferred.
  • the output can be qualitative (eg, “positive” or “negative”) or quantitative (eg, a concentration such as nanograms per milliliter).
  • the present invention includes a method for detecting pancreatic cancer, comprising detecting and / or quantifying the presence of the biomarker of the present invention in a body fluid sample obtained from a subject.
  • the method of the invention further comprises comparing the level of the biomarker of the invention in the body fluid sample with the level of the biomarker of the invention present in the normal control sample, wherein the level in the normal control sample Compared with, a high level or low level of the biomarker of the present invention in a body fluid sample is an indicator of the presence of pancreatic cancer.
  • the invention comprises detecting and / or measuring the presence of a biomarker of the invention in a body fluid sample obtained from a subject such as urine, blood, etc.
  • a method of detecting cancer in a subject that is indicative of the presence of cancer in the subject when the level of the marker is higher / lower than a predetermined control level. Detection is preferably performed by immunological methods or mass spectrometry.
  • the level of the biomarker of the present invention detected in the body fluid sample obtained from the subject is higher than the level of the corresponding biomarker in the control sample.
  • the sample obtained from the subject AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6, ATP6AP1
  • the biomarker level is higher than the level of the corresponding biomarker in the control sample.
  • the level of the biomarker of the present invention detected in the body fluid sample obtained from the subject is lower than the level of the corresponding biomarker in the control sample.
  • HYOU1 in the subject sample The level is lower than the control level.
  • the biomarker of the present invention may be used alone, or may be used as a panel combined with a known biomarker or a panel combined with the biomarker of the present invention.
  • a plurality of biomarkers By using a plurality of biomarkers, an improvement in detection sensitivity of pancreatic cancer can be expected.
  • the combination of biomarkers is not particularly limited, and those skilled in the art can appropriately select the biomarkers in consideration of ease of detection, certainty, high detection sensitivity, and the like.
  • Standards or controls used in the methods of the present invention can be obtained from samples of biomarker levels of the present invention obtained from healthy control subjects, samples obtained from subjects with benign diseases (benign tumors), or other tests. It may correspond to a biomarker level of the present invention obtained from a body sample. An increase or decrease in the level of a biomarker of the present invention compared to a standard may indicate the presence of early to late stage pancreatic cancer.
  • the present invention provides: a) measuring the level of a biomarker of the present invention in a body fluid sample such as urine, blood, etc. obtained from a subject; and b) the level measured in step (a). Comparing to a range of biomarkers of the invention known to be present in a body fluid sample obtained from a normal subject not having cancer, and c) testing based on the comparison of step (b) Determining the susceptibility of the body to pancreatic cancer, wherein the high level or low level of the biomarker of the present invention in step (a) determines the susceptibility to precancerous lesions or risk diseases. Includes methods to evaluate.
  • the method of the present invention is suitable for diagnosing and monitoring pancreatic cancer by quantifying the biomarker of the present invention in a body fluid sample obtained from a subject.
  • the amount of a biomarker of the invention quantified in a sample obtained from the subject being tested is a level quantified for another sample or a previous sample obtained from the subject, or a level quantified for a control sample It is preferable to compare.
  • healthy subjects with no clinically apparent disease or abnormality can be selected as controls.
  • Diagnosis can be made by finding a statistically different level of a biomarker of the invention compared to a previous level quantified for a control sample or the same subject.
  • the present invention provides a method for diagnosis and monitoring of pancreatic cancer in a subject by detecting the biomarker of the present invention in a body fluid sample obtained from the subject.
  • the method comprises contacting a sample with an antibody specific for a biomarker of the invention that is directly or indirectly labeled with a detectable substance and detecting the detectable substance.
  • the method includes detecting the level of a biomarker of the invention using a mass spectrometer.
  • the method of the present invention is useful for diagnosis of early pancreatic cancer (for example, when the subject is asymptomatic) and for prognosis of pancreatic cancer progression and death.
  • a biomarker of the invention detected in a body fluid sample eg, urine, serum, plasma, whole blood
  • a standard eg, normal or benign disease level
  • a decline indicates an increased disease stage, disease progression or increased risk of death.
  • the subject suffers from pancreatic cancer and detection is performed at several time points at intervals as part of subject monitoring before, during or after cancer treatment.
  • the subject does not show symptoms of cancer when the detection of the biomarkers of the invention is performed. In other embodiments, the subject exhibits one or more cancer symptoms at the time detection of the biomarkers of the invention is performed.
  • the method of the invention detects a cancer biomarker in the same body fluid sample or a different body fluid sample obtained from a subject before, during or after the detection of the biomarker of the invention described above.
  • the biomarker for pancreatic cancer is CA19-9.
  • the subject may have an elevated level of CA19-9 in the blood when the detection of the biomarker of the present invention is performed, or the blood may be detected when the detection of the biomarker of the present invention is performed. In CA19-9, the level may not have risen.
  • the biomarker of the present invention in a body fluid sample of a subject can be detected by a drug that interacts or binds to the biomarker protein of the present invention or a fragment thereof.
  • agents include the biomarker antibodies of the invention or fragments thereof that bind to the biomarkers of the invention. That is, the present invention also includes a method for detecting an antibody against the biomarker of the present invention.
  • the biomarker of the present invention is a marker for pancreatic cancer.
  • detection of a biomarker antibody of the present invention in a body fluid of a subject can enable diagnosis of pancreatic cancer.
  • An antibody that binds to the biomarker of the present invention can be used as a marker for pancreatic cancer.
  • antibody refers to immunoglobulin molecules and immunologically active portions (fragments) of immunoglobulin molecules, ie, molecules comprising an antibody binding site or paratope.
  • Antibodies include monoclonal antibodies, polyclonal antibodies, and antigen binding fragments thereof.
  • the antibody of the present invention may be labeled with a detectable substance (for example, a detectable moiety).
  • antibodies As the antibody specific for the biomarker of the present invention used in the method of the present invention, a commercially available antibody can be used.
  • antibodies, monoclonal or polyclonal antibodies, and immunologically active fragments eg, Fab or (Fab) 2 fragments
  • antibody heavy chains eg, Fab or (Fab) 2 fragments
  • antibody light chains e.g., humanized antibodies
  • genetically engineered single chain Fv molecules or chimeric antibodies e.g, Fab or (Fab) 2 fragments
  • Antibodies such as monoclonal and polyclonal antibodies, fragments and chimeras, can be prepared using methods known to those skilled in the art.
  • the detection comprises contacting a body fluid sample from a subject with an antibody that binds to a biomarker protein of the invention, and at least one biomarker in the body fluid sample and the antibody. Detecting and / or quantifying the binding to.
  • One embodiment of the invention also includes correlating the detected binding with the amount of the biomarker protein of the invention in a sample.
  • the method further comprises the step of comparing the level of the biomarker in the body fluid sample with the control level of the biomarker, wherein the biomarker level in the subject sample is higher or lower than the control level. In some cases, it is an indicator that pancreatic cancer is present.
  • the invention comprises (a) contacting a body fluid sample from a subject with an antibody specific for a biomarker of the invention that is directly or indirectly labeled with a detectable substance (reaction). And (b) detecting a detectable substance, and a method for detecting pancreatic cancer in a subject, comprising quantifying the biomarker of the present invention in the body fluid sample.
  • the invention comprises (a) contacting a body fluid sample from a subject with an antibody specific for a biomarker of the invention that is directly or indirectly labeled with a detectable substance (reaction). Diagnosing and / or monitoring pancreatic cancer in a subject comprising quantifying a biomarker of the present invention in said bodily fluid sample, comprising: (b) detecting a detectable substance Regarding the method.
  • An embodiment of the method of the invention comprises (a) contacting a bodily fluid sample from a subject with an antibody specific for a biomarker of the invention that is directly or indirectly labeled with an enzyme, and (b) Adding a substrate of the enzyme, wherein the substrate is selected such that the substrate or the reaction product of the enzyme and the substrate forms a fluorescent complex, and (c) measuring the fluorescence of the fluorescent complex Quantifying the biomarker of the invention in and (d) comparing the quantified level to that of a standard.
  • a preferred embodiment of the present invention includes the following steps: (A) A body fluid sample together with a primary antibody specific for the biomarker of the present invention that is directly or indirectly labeled with a detectable substance and a secondary antibody specific for the biomarker of the present invention that is immobilized Incubating, (B) separating the primary antibody from the secondary antibody to provide a primary antibody phase and a secondary antibody phase; (C) detecting a detectable substance in the primary or secondary antibody phase, thereby quantifying the biomarker of the invention in a body fluid sample; (D) comparing the quantified biomarker of the invention to a standard.
  • the present invention provides a method for measuring the biomarker of the present invention in a body fluid sample by measuring the biomarker of the present invention by an immunological assay according to the embodiment. It will be apparent to those skilled in the art that various immunological assays can be used to measure the biomarkers of the present invention. In general, the immunological assays used in the present invention may be competitive or non-competitive.
  • the competition method usually uses an immobilized or immobilizable antibody of the biomarker of the present invention (an anti-biomarker of the present invention) and a labeled form of the biomarker of the present invention.
  • the biomarker of the present invention and the labeled biomarker of the present invention in the sample compete for binding to the antibody.
  • labeling in either the bound or unbound fraction can be correlated with the amount of the biomarker of the invention in a body fluid sample by any conventional method, for example by comparison with a standard curve.
  • the biomarker of the present invention in a body fluid sample can be detected using an antibody or derivative that specifically reacts with the biomarker of the present invention, such as an enzyme conjugate or a labeled derivative.
  • an antibody or derivative that specifically reacts with the biomarker of the present invention such as an enzyme conjugate or a labeled derivative.
  • they can be used in any known immunological assay that relies on binding interactions between antigenic determinants of proteins and antibodies.
  • the immunological assay method used in the present invention may be any method that can be performed by those skilled in the art, and the above description is an example of the assay method and does not limit the scope of the present invention.
  • immunological assays include Western blot, dot blot analysis, slot blot analysis, radioimmunoassay (RIA), peptide microarray, enzyme immunoassay (EIA) and enzyme linked immunosorbent assay (ELISA), There are immunofluorescence, immunoprecipitation, latex aggregation, hemagglutination and histochemical examination.
  • ELISA refers to an antibody bound to a solid phase in order to detect and / or quantify the amount of an antigen (eg, a biomarker of the present invention) or antibody present in a sample.
  • ELISA can be performed by binding an antibody specific for the antigen or protein of interest on a solid support (eg, polyvinyl chloride).
  • a solid support eg, polyvinyl chloride
  • the cell extract or other sample of interest such as urine, can be added and the excess unbound sample is washed away.
  • Add an enzyme-bound antibody specific for a different site on the antigen The support is washed to remove secondary antibody bound by unbound enzyme. Examples of the antibody to which the enzyme is bound include alkaline phosphatase.
  • Enzymes on the secondary antibody can convert the added colorless substrate into a colored product or convert a non-fluorescent substrate into a fluorescent product.
  • the antibody specific for the biomarker of the present invention can be labeled with a detectable substance, and the position in the body fluid sample can be specified based on the presence of the detectable substance.
  • label and “tag” refer to substances that can provide a detectable signal, including but not limited to enzymes such as alkaline phosphatase, glucose-6-phosphate dehydration.
  • Elementary enzymes and replicase substrates such as horseradish peroxidase, ribozyme, QB replicase, promoters, dyes, fluorescent substances such as fluorescein, isothiocyanate, rhodamine compounds, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine, chemiluminescent substances, Examples include isoluminol, sensitizers, coenzymes, enzyme substrates, radiolabels, particles such as latex or carbon particles, liposomes, cells, etc., which are further treated with dyes, catalysts or other detectable groups. It can be labeled.
  • An indirect method in which the primary antigen-antibody reaction is amplified by the introduction of a secondary antibody having specificity for an antibody reactive with the biomarker of the present invention can also be used.
  • the antibody having specificity for the biomarker of the invention is a rabbit IgG antibody
  • the secondary antibody is a goat anti-rabbit gamma globulin labeled with a detectable substance described herein. obtain.
  • the antibody is either shared directly or via a linker with a compound that serves as a reporter group to allow detection of the presence of the biomarkers of the invention.
  • a linker with a compound that serves as a reporter group to allow detection of the presence of the biomarkers of the invention.
  • a variety of different types of substances can serve as reporter groups, including but not limited to enzymes, dyes, radioactive and non-metallic isotopes, fluorescent compounds, fluorescent compounds, and the like.
  • a method for conjugating or labeling an antibody can be easily achieved by those skilled in the art.
  • an enzyme substrate selected such that the biomarker antibody of the present invention is labeled with an enzyme, and the substrate or the reaction product of the enzyme and the substrate forms a fluorescent complex with the lanthanide metal. Provides an added method.
  • the biomarker of the present invention in the sample is quantified by adding lanthanide metal and measuring the fluorescence of the fluorescent complex.
  • Antibodies specific for the biomarkers of the present invention can be labeled directly or indirectly using enzymes.
  • the enzyme is selected based on the ability of the enzyme substrate or enzyme-substrate reaction product to form a complex with lanthanide metals such as europium and terbium.
  • suitable enzymes include alkaline phosphatase and ⁇ -galactosidase.
  • the enzyme is preferably alkaline phosphatase.
  • the biomarker antibody of the present invention can also be indirectly labeled with an enzyme.
  • the antibody can be conjugated with one partner of the ligand binding pair and the enzyme can be coupled with the other partner of the ligand binding pair.
  • Representative examples include avidin-biotin and riboflavin-riboflavin binding protein.
  • the antibody is biotinylated and the enzyme is coupled with streptavidin.
  • an antibody bound to a biomarker of the present invention in a sample is detected by adding an enzyme substrate.
  • the substrate is selected such that the substrate or enzyme-substrate reaction product forms a fluorescent complex with the lanthanide metal in the presence of a lanthanide metal (eg, europium, terbium, samarium and dysprosium, preferably europium and terbium).
  • a lanthanide metal eg, europium, terbium, samarium and dysprosium, preferably europium and terbium.
  • the substrate used in the method may be 4-methylumbelliferyl phosphate or 5-fluorosalicyl phosphate.
  • the fluorescence intensity of the complex is usually measured using a time-resolved fluorometer, such as a CyberFluor 615 Immunoanalyzer (Nordion International, Ontario, Canada).
  • the sample, an antibody specific for the biomarker of the present invention, or the biomarker of the present invention may be immobilized on a carrier.
  • suitable carriers include agarose, cellulose, dextran, sephadex, sepharose, liposomes, carboxymethylcellulose polystyrene, filter paper, ion exchange resins, plastic films, plastic tubes, glass beads, polyamine-methyl vinyl ether-maleic acid copolymers, Examples include amino acid copolymers, ethylene-maleic acid copolymers, nylon, and silk.
  • the carrier may be in the form of, for example, a tube, a test plate, a well, a bead, a disk, a sphere, and the like.
  • Immobilized antibodies can be prepared by reacting the material with a suitable insoluble carrier using known chemical or physical methods such as cyanogen bromide coupling.
  • biomarker antibodies of the invention are used.
  • One of the biomarker antibodies of the invention is directly or indirectly labeled (also referred to as “detection antibody”), and the other is immobilized or can be immobilized (also referred to as “capture antibody”).
  • Capture and detection antibodies can be contacted simultaneously or sequentially with the body fluid sample.
  • Sequential methods can be accomplished by incubating the capture antibody with the sample and then adding the detection antibody at a predetermined time (sometimes referred to as the “forward method”), or first, incubating the detection antibody with the sample, A capture antibody can then be added (sometimes referred to as the “reverse” method). After performing the incubation or the like necessary to complete the assay, the capture antibody is separated from the liquid test mixture and the label in at least a portion of the separated capture antibody phase or the remainder of the liquid test mixture is measured. Usually measured in the capture antibody phase because it contains a biomarker of the invention bound by and “sandwiched” between the capture and detection antibodies.
  • one or both of the capture antibody and the detection antibody are polyclonal antibodies.
  • the label used for the detection antibody can be selected from any of those conventionally known in the art.
  • the label is detected by, for example, secondary binding by an enzyme or chemiluminescent moiety or radioisotope, a fluorophore, a detectable ligand (eg, a labeled binding partner of the ligand). Possible).
  • the antibody is preferably labeled with an enzyme that is detected by the addition of a substrate that is selected so that the reaction product of the enzyme and substrate forms a fluorescent complex.
  • the capture antibody is selected to provide a method that is separated from the remainder of the test mixture.
  • the capture antibody may be already immobilized or introduced into the assay in an insoluble form, or in an immobilizable form, i.e., immobilization that is subsequently achieved to introduce the capture antibody into the assay. It may be in a form that enables.
  • Immobilized capture antibodies can include antibodies that are covalently or non-covalently bound to a solid phase such as magnetic particles, latex particles, microtiter multiwell plates, beads, cuvettes or other reaction vessels. .
  • immobilizable capture antibodies are chemically modified with a ligand moiety, eg, hapten, biotin, etc., followed by an immobilized form of a ligand binding partner, eg, antibody, avidin, etc.
  • a ligand moiety eg, hapten, biotin, etc.
  • an immobilized form of a ligand binding partner eg, antibody, avidin, etc.
  • the capture antibody can be immobilized using a species-specific antibody of the capture antibody that is bound to the solid phase.
  • the specific sandwich immunoassay method of the present invention comprises two antibodies reactive to the biomarker of the present invention, an antibody reactive to the biomarker of the present invention labeled with an enzyme label.
  • a secondary antibody having specificity for the enzyme and a fluorescent substrate of the enzyme are used.
  • the enzyme is alkaline phosphatase (ALP) and the substrate is 5-fluorosalicyl phosphate.
  • ALP cleaves phosphate from a fluorescent substrate, 5-fluorosalicyl phosphate, to produce 5-fluorosalicylic acid (FSA).
  • 5-Fluorosalicylic acid then forms a highly fluorescent triple complex in the form of FSA-Tb (3 +)-EDTA, which can be quantified by measuring Tb 3+ fluorescence in a time-resolved manner. Fluorescence intensity is usually measured using a time-resolved fluorometer as described herein.
  • the method of the present invention can be carried out on a solid support.
  • the solid support used may be conventional for the purpose of assaying an analyte in a body fluid sample and is usually composed of materials such as cellulose, polysaccharides, eg Sephadex, etc. And / or may be partially surrounded by a covering for handling.
  • the solid support may be rigid, semi-rigid, flexible, elastic (having shape memory), etc., depending on the desired application.
  • the biomarker of the present invention can be detected in a sample in vivo or in vitro (ex vivo).
  • the support is harmless to the subject. It must be in any form convenient for insertion into an appropriate part of the body.
  • the support can be a probe made of polytetrafluoroethylene, polystyrene or other rigid non-hazardous plastic material and having a size and shape that allows it to be introduced into a subject.
  • the selection of a suitable inert support is within the ability of one skilled in the art, such as dimensions for the intended purpose.
  • the contacting step in the method of the present invention comprises contacting or combining a body fluid sample and a solid support, for example, a reaction vessel, micro container, test tube, micro test tube, well, multi-well plate or other solid support. Or may include mixing.
  • a solid support for example, a reaction vessel, micro container, test tube, micro test tube, well, multi-well plate or other solid support. Or may include mixing.
  • the sample and / or the biomarker specific binding agent of the present invention may be disposed on a solid support, or a plurality of supports may be used.
  • “Sequencing” refers to library members (eg, arrays of different samples or arrays of devices that target the same or different target molecules) or other collections, logical arrays or physical Refers to the act of organizing or arranging in an array.
  • a biomarker in a body fluid sample derived from a subject can be detected and / or measured using a mass spectrometer.
  • a mass spectrometer not only the mass of a peptide eluted from HPLC, but also fragmented peptides of various lengths generated by cleaving the peptide by giving excessive energy to the peptide when ionizing the peptide ( The mass of the cleaved peptide) can also be measured.
  • the amino acid sequence which comprises a peptide can be determined by analyzing the difference in mass of these cleavage peptides.
  • a specific peptide having a known amino acid sequence is determined from a mixture of peptides having many different masses, and the mass of the amino acid sequence and the mass of a cleaved peptide that can be generated from the peptide. It can be detected using. That is, using the mass predicted from the amino acid sequence of the tryptic peptide fragment of at least one biomarker and the mass of the cleaved peptide predicted from the peptide, at least one in a blood sample from a pancreatic cancer patient It can be confirmed whether or not there is a molecule having a sequence unique to the biomarker.
  • a commercially available apparatus can be used as the mass spectrometer, and those skilled in the art can appropriately carry out the method of the present invention using a mass spectrometer using a known technique.
  • kits and diagnostic agents for diagnosing or monitoring cancer includes kits comprising elements necessary for detecting, diagnosing or monitoring pancreatic cancer.
  • the kit includes an agent (eg, an antibody) for detecting the presence of the biomarker of the present invention in a patient-derived body fluid.
  • the antibody against the biomarker of the present invention may preferably include a container for collecting body fluid from a patient.
  • the antibody included in the kit may be in the form of an aqueous medium or a lyophilized product.
  • kits or diagnostic agents for qualitatively or quantitatively detecting the biomarker of the present invention in a sample such as blood or urine.
  • a kit or diagnostic agent can include a binding agent (eg, an antibody) specific for a biomarker of the invention, an antibody to an antibody labeled with an enzyme, and an enzyme substrate.
  • the kit or diagnostic agent can also be used to carry out the methods of the invention using a solid support such as a microtiter multiwell plate, standard, assay diluent, wash buffer, adhesive plate cover and / or kit. Instructions for use may be included.
  • the kit or diagnostic agent may include one or more protease inhibitors (eg, a protease inhibitor cocktail) that are applied to a body fluid sample (eg, blood or urine) to be assayed.
  • the kit or diagnostic agent of the present invention includes, for example, means for collecting a body fluid sample, means for labeling a detection agent (binding agent), the biomarker protein of the present invention in the body fluid sample, or the biomarker nucleic acid of the present invention.
  • a detection agent binding agent
  • Membrane means for applying a body fluid sample to the membrane, means for binding the agent to the biomarker of the present invention in the body fluid sample of the subject, secondary antibodies and the like.
  • the present invention can use conventional techniques in molecular biology, microbiology, recombinant DNA technology, electrophysiology, and pharmacology, which are within the scope of those skilled in the art.
  • Such techniques are well described in the literature (eg, Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory Manual, No. 1). 2nd edition (1989); DNA Cloning, Volumes I and II (DN Glover, 1985); Perval, B., A Practical Guide to Two Molecular Cloning (A Practical Guide to Molecular Cloning) (1984); The Series, Methods in Enzymology (the) eries, Methods In Enzymology (S. Colowick and N.
  • the plasma microarray (Murakoshi Y, Nissan K, Sasakiki S, Ono M, Negishi A, Matsubara J, Sakutama T, Kuvabara H, Nakamori S, Nakamori S, originally developed at the National Cancer Center. Ioka T, Okusaka T, Kosuge T, Shimahara M, Yasunami Y, Ino Y, Tsuchida A, Aoki T, Tsugane S, Yamada T.Plasma biomarker discovery and validation for colorectal cancer by quantitative shotgun mass spectrome . Ry and protein microarray.Cancer Sci.2011 Mar; 102 (3): 630-8, also referred to as "Murakoshi et al”) was subjected to hybridization at 130 antibody panel was measured protein expression in the plasma of each antigen.
  • the plasma microarray used for measuring each antigen amount according to the present example was prepared by the same method as Murakoshi et al. Antibody hybridization, fluorescence intensity measurement, and the like were also carried out in the same manner as Murakoshi et al. DNA / antibody / protein microarrayer (KAKEN GENEX) was used for the production of plasma microarray, and Innoscan 700AL (Inopsys) was used for the measurement of fluorescence intensity.
  • KAKEN GENEX DNA / antibody / protein microarrayer
  • Innoscan 700AL Inopsys
  • the plasma microarray used in this example includes 106 healthy subjects, 164 cases of invasive pancreatic duct cancer, 7 cases of pancreatic malignant tumor other than invasive pancreatic duct cancer, 15 cases of benign pancreatic tumor / cyst, 10 cases of chronic pancreatitis, 11 cases of hepatocellular carcinoma, 13 cases of biliary tract cancer, 30 cases of gastric cancer. 28 cases of colorectal cancer were mounted.
  • Biomarker identification Protein expression was digitized as fluorescence intensity, and 23 types of antibodies that discriminate between pancreatic cancer patients and healthy individuals with an AUC value of 0.8 or higher were selected.
  • the fluorescence intensity of the 23 types of antibodies increased or decreased with a statistically significant difference between pancreatic cancer patients and healthy individuals (Table 4).
  • 23 types of antigen proteins AP3B1, C2, CKS1B, CKS2, CSPG2, CYCS, CD82, PI3, RNASE1, RNASET2, VRK2, EV1, HMGB2, MST4, MMP9, MYBL2, PPM1B, AK3L1, IGFBP2, STMN1, ANXA6 ATP6AP1 was statistically significantly increased in the serum of patients with invasive pancreatic duct cancer compared to healthy subjects.
  • HYOU1 antigen was significantly decreased in patients with invasive pancreatic duct cancer compared to healthy subjects (FIGS. 1 to 23).
  • the case data shown in FIGS. 1 to 23 will be described.
  • the table on the upper left of the figure shows the antibody name, ID number, plate T (dilution ratio of plasma), test P value (p- between healthy subjects and patients with invasive pancreatic duct cancer when calculated at each concentration) from the left. value (Student's t-test)), AUC (area under the curve when ROC analysis is performed), threshold. 2SD (corresponding to the value obtained by adding a value twice the standard deviation to the average value of healthy subjects, corresponding to the Cut-off value), sensitivity SD. 2SD (threshold. Sensitivity when 2SD is cut-off), specificity. 2SD (threshold. Sensitivity when 2SD is cut-off) is shown.
  • the leftmost figure in the middle row shows a box-and-whisker plot of the fluorescence values of the plasma microarray for healthy subjects and each disease.
  • 1 106 healthy subjects
  • 2 164 cases of invasive pancreatic duct cancer
  • 3 7 cases of pancreatic malignant tumor other than invasive pancreatic duct cancer
  • 4 15 cases of benign pancreatic tumor / cyst
  • 5 10 cases of chronic pancreatitis
  • 6 Results of 11 cases of hepatocellular carcinoma
  • 8 30 cases of gastric cancer
  • 9 28 cases of colorectal cancer.
  • the figure in the middle row shows a scatter diagram of the fluorescence values of the plasma microarray for each healthy subject and each disease.
  • 1 106 healthy subjects
  • 2 164 cases of invasive pancreatic duct cancer
  • 3 7 cases of pancreatic malignant tumor other than invasive pancreatic duct cancer
  • 4 15 cases of benign pancreatic tumor / cyst
  • 5 10 cases of chronic pancreatitis
  • 6 Results of 11 cases of hepatocellular carcinoma
  • 8 30 cases of gastric cancer
  • 9 28 cases of colorectal cancer.
  • the leftmost figure at the bottom is a scatter diagram when CA19-9 is combined with a marker.
  • the X axis shows the measured value (Unit) of CA19-9, and the Y axis shows the fluorescence value (Unit) of the marker.
  • the figure in the lower row shows an ROC curve of a marker for discriminating healthy subjects from invasive pancreatic duct cancer.
  • the X axis indicates 1-specificity, and the Y axis indicates sensitivity.
  • the right figure shows the fluorescence scanner image when the plasma microarray was hybridized with the marker antibody.
  • Table 6 shows the AUC values and correlation coefficients when the biomarkers of the present invention are combined.
  • a high AUC value was obtained by combining a plurality of, for example, two biomarkers of the present invention compared to a single case (Tables 4 and 5).
  • a significant AUC value was obtained for both the combination of the biomarker of the present invention and a known biomarker, and the combination of the biomarkers of the present invention. From these results, it was shown that the detection sensitivity of pancreatic cancer can be further improved by using a plurality of the biomarkers of the present invention.
  • the present invention provides a biomarker for detecting pancreatic cancer, a method for detecting pancreatic cancer using the biomarker, and a kit or diagnostic agent for detecting pancreatic cancer containing an antibody against the biomarker.
  • the biomarker of the present invention can be used for efficiently discriminating pancreatic cancer, and a method for detecting or diagnosing pancreatic cancer using these biomarkers is extremely effective clinically.

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Abstract

L'invention concerne un procédé de détection du cancer du pancréas, ledit procédé consistant à détecter et/ou à doser une protéine ou un fragment de cette dernière existant dans un échantillon de fluide corporel d'un sujet. L'invention concerne également un agent de diagnostic pour le cancer du pancréas ou un kit de détection du cancer du pancréas, comprenant chacun un anticorps contre la protéine susmentionnée ou un fragment de cette dernière.
PCT/JP2016/066514 2015-05-29 2016-05-27 Panel de biomarqueurs de plasma permettant le diagnostic du cancer du pancréas WO2016195051A1 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111323589A (zh) * 2020-03-03 2020-06-23 南通大学附属医院 血清外泌体anxa11在诊断胰腺癌中的应用
WO2021006649A1 (fr) * 2019-07-09 2021-01-14 주식회사 베르티스 Panel de biomarqueurs pour le diagnostic du cancer du pancréas et utilisation de celui-ci
WO2023282916A1 (fr) 2021-07-09 2023-01-12 Guardant Health, Inc. Procédés de détection de réarrangements génomiques à l'aide d'acides nucléiques acellulaires
EP4049027A4 (fr) * 2019-10-24 2024-02-07 The Council of the Queensland Institute of Medical Research Diagnostic du cancer
EP4407042A2 (fr) 2020-07-10 2024-07-31 Guardant Health, Inc. Procédés de détection de réarrangements génomiques à l'aide d'acides nucléiques acellulaires

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JP2006500948A (ja) * 2002-09-30 2006-01-12 オンコセラピー・サイエンス株式会社 膵臓癌を診断する方法
JP2010175452A (ja) * 2009-01-30 2010-08-12 Japan Health Science Foundation 膵臓癌の検出法
JP2013027387A (ja) * 2011-06-21 2013-02-07 Tohoku Univ 膵臓癌バイオマーカー

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
JP2006500948A (ja) * 2002-09-30 2006-01-12 オンコセラピー・サイエンス株式会社 膵臓癌を診断する方法
JP2010175452A (ja) * 2009-01-30 2010-08-12 Japan Health Science Foundation 膵臓癌の検出法
JP2013027387A (ja) * 2011-06-21 2013-02-07 Tohoku Univ 膵臓癌バイオマーカー

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021006649A1 (fr) * 2019-07-09 2021-01-14 주식회사 베르티스 Panel de biomarqueurs pour le diagnostic du cancer du pancréas et utilisation de celui-ci
EP4049027A4 (fr) * 2019-10-24 2024-02-07 The Council of the Queensland Institute of Medical Research Diagnostic du cancer
CN111323589A (zh) * 2020-03-03 2020-06-23 南通大学附属医院 血清外泌体anxa11在诊断胰腺癌中的应用
EP4407042A2 (fr) 2020-07-10 2024-07-31 Guardant Health, Inc. Procédés de détection de réarrangements génomiques à l'aide d'acides nucléiques acellulaires
WO2023282916A1 (fr) 2021-07-09 2023-01-12 Guardant Health, Inc. Procédés de détection de réarrangements génomiques à l'aide d'acides nucléiques acellulaires

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