WO2017054686A1 - Produit, application et procédé utilisés pour permettre une évaluation et un criblage se rapportant à des tumeurs malignes - Google Patents

Produit, application et procédé utilisés pour permettre une évaluation et un criblage se rapportant à des tumeurs malignes Download PDF

Info

Publication number
WO2017054686A1
WO2017054686A1 PCT/CN2016/100048 CN2016100048W WO2017054686A1 WO 2017054686 A1 WO2017054686 A1 WO 2017054686A1 CN 2016100048 W CN2016100048 W CN 2016100048W WO 2017054686 A1 WO2017054686 A1 WO 2017054686A1
Authority
WO
WIPO (PCT)
Prior art keywords
gal
ratio
subject
cancer
galactose
Prior art date
Application number
PCT/CN2016/100048
Other languages
English (en)
Chinese (zh)
Inventor
任士芳
顾建新
周蕾
孙琳琳
Original Assignee
上海知先生物科技有限公司
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 上海知先生物科技有限公司 filed Critical 上海知先生物科技有限公司
Publication of WO2017054686A1 publication Critical patent/WO2017054686A1/fr

Links

Images

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
    • G01N33/50—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
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6848—Methods of protein analysis involving mass spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6848—Methods of protein analysis involving mass spectrometry
    • G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins

Definitions

  • the invention belongs to the fields of biotechnology and medicine.
  • the present invention relates to screening, early diagnosis, prognostic evaluation, risk assessment, condition monitoring and/or monitoring of malignant tumors by detecting changes in the immunoglobulin G surface dual-antenna complex N-glycan terminal galactose in the blood. Products and methods for evaluation of efficacy.
  • Tumor is one of the diseases that threaten human health in the world today. It has long-term and multi-stage characteristics. When a patient has a self-conscious symptom to the hospital, the tumor often develops into the middle and late stage, which brings great difficulties to clinical treatment. The mortality rate of advanced tumors is very high, and the five-year survival rate is often only 20%. If the tumor can be detected early and monitored for development and timely treatment, the five-year survival rate of the tumor can reach 80%. At present, the medical community generally believes that secondary prevention (early detection, early diagnosis, early treatment) is one of the key links to reduce cancer mortality. It is worth mentioning that, from the age incidence rate, the incidence rate increased significantly after the age of 45. Therefore, screening for malignant tumors from the age of 40 has great social significance.
  • screening methods for malignant tumors mainly include imaging examination and screening of serum tumor markers.
  • imaging examinations including CT, nuclear magnetic resonance, endoscopy, etc.
  • these conventional imaging examination methods It is to identify the tissue morphology, and often only detect the middle and late stage tumors of a certain size.
  • serological tumor detection methods are more suitable for extensive tumor screening.
  • serum tumor markers including alpha-fetoprotein (AFP), cancer antigen 125 (CA125), carcinoembryonic antigen (CEA), cancer antigen 15-3 (CA15-3), and prostate specific antigen (PSA).
  • AFP alpha-fetoprotein
  • CA125 cancer antigen 125
  • CEA carcinoembryonic antigen
  • CA15-3 cancer antigen 15-3
  • PSA prostate specific antigen
  • these tumor markers are basically directed to a certain type or a certain type of tumor, which is difficult to comprehensively consider when used for testing.
  • the prognosis of malignant tumors and the assessment of the risk of malignant tumors in high-risk populations, patient condition monitoring and / or efficacy evaluation are also important for the clinical control of malignant tumors, recurrence, treatment and so on.
  • sugar chains are involved in almost all life processes.
  • the changes in sugar chain structure are closely related to the occurrence and development of diseases such as tumors, autoimmune diseases and infections.
  • diseases such as tumors, autoimmune diseases and infections.
  • some glycoproteins on the cell membrane have been abnormally glycosylated and released into the blood, which has become an important clue for early detection of tumors.
  • a variety of tumor serodiagnosis biomarkers such as CA19-9 (digestive tract tumor), AFP (liver cancer), CA125 (ovarian cancer) are all sugar chain or glycoprotein products secreted by the body after tumorigenesis.
  • the in-depth study based on the sugar field provides an opportunity for the development of new hematological tumor detection methods.
  • the inventors have found that the degree of terminal galactosylation of a double-antenna complex N-glycan chain on human serum IgG (specifically, N-glycochain end-galactose (Gal 0 ) and galactose chain (Gal 1 and Gal 2 ) abundance Ratio) has a general correlation with the occurrence, benign and malignant, and prognosis of various tumors. Based on this, the present invention provides novel methods and products for malignant tumor screening, prognosis assessment, risk assessment, condition monitoring, and/or efficacy assessment.
  • a product for screening, early diagnosis, prognostic evaluation, risk assessment, condition monitoring and/or efficacy evaluation of a subject which may be, for example, a kit, a device , an operating system, and/or a combination thereof, the product comprising:
  • the sugar chain abundance Gal 0 the double-antenna complex N-glycan abundance Gal 1 with a galactose at the end and the double antenna complex N-glycan abundance Gal 2 with 2 galactose at the end;
  • reagents, instruments, and/or systems for one or more methods selected from the group consisting of matrix-assisted laser desorption time-of-flight mass spectrometry MALDI-MS, fast atom bombardment mass spectrometry FAB-MS, electrospray mass spectrometry ESI-MS ; liquid chromatography; liquid chromatography-mass spectrometry; sugar chip technology; nuclear magnetic resonance NMR, preferably matrix-assisted laser desorption time-of-flight mass spectrometry MALDI method;
  • the blood is selected from the group consisting of serum, plasma, and whole blood, preferably serum.
  • the product further comprises one or more selected from the group consisting of:
  • reagents and/or instruments for collecting and/or processing a blood sample of a subject
  • e a database, module and/or processor for the ratio of the double-antennary complex N-glycan end galactose to galactose chain abundance for storing and/or processing the subject serum as an indicator of galactose granulation of the IgG surface;
  • a module and/or processor for comparing a Gal ratio of a subject to a control to determine whether the subject has a malignancy, a prognostic assessment of the subject, and/or a risk assessment, condition monitoring, and/or efficacy assessment of the subject;
  • modules and/or processors for providing diagnostic and/or test results and/or reports
  • (v) for cancer patient condition monitoring when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a predetermined range (for example, 5% to 50% or any point or sub-range within the range), It indicates that the condition of the cancer patient has developed or worsened;
  • a predetermined range for example, 5% to 50% or any point or sub-range within the range
  • Gal ratios are measured and calculated at different time points before and after treatment or during treatment, when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a preset range (eg 5%) ⁇ 50% or any point or sub-range within the range) indicates that the therapy and/or drug is not effective;
  • the threshold value in (i) to (iv) is 0.35 to 0.6, and more preferably 0.5.
  • the maximum slope or maximum sensitivity and specificity in the ROC curve can be based
  • the value is determined to determine the optimal threshold, and the preferred threshold range may be the range of the optimal threshold ⁇ 1 to 15%, including any numerical or sub-range within the range.
  • control is selected from the group consisting of a sample obtained from a healthy or non-malignant tumor patient, or a predetermined threshold.
  • the values of a, b, and c are independently between 0 and 10, and a ⁇ 0, b and c are not 0 at the same time.
  • a, b, and c are each independently selected from the group consisting of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.2, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9 or 10.
  • the specific values of a, b or c can be adjusted depending on the degree of response of the various parameters Gal 0 , Gal 1 and Gal 2 in the test instrument and/or method used. For example, the specific values of a, b or c can be adjusted based on the ratio of the test method used to the standard method (eg, ultra high performance liquid chromatography, UPLC).
  • the standard method eg, ultra high performance liquid chromatography, UPLC
  • a ratio calculation can be performed using a formula selected from the group consisting of:
  • Gal ratio Gal 0 / (Gal 1 + Gal 2 );
  • Gal ratio Gal 0 / (Gal 1 + 2Gal 2 ); or
  • the malignant tumor is selected from the group consisting of liver cancer, gastric cancer, lung cancer, ovarian cancer, intestinal cancer, esophageal cancer, pancreatic cancer, cholangiocarcinoma, renal cancer, breast cancer, prostate cancer, nasopharyngeal cancer, Bladder cancer, endometrial cancer and cervical cancer.
  • the malignant tumor is in a stage selected from the group consisting of precancerous lesions, preclinical cancer, early cancer, and advanced cancer.
  • the subject is a mammal, preferably a human, a monkey, a dog, a horse, a cow, a sheep, a pig, a mouse, a rabbit, etc., more preferably a high risk population of cancer.
  • the product is for high throughput detection, such as for simultaneous processing of 96, 192, 288, 384, 480, 576 or more samples.
  • the product is used in a detection method comprising the following steps:
  • the sugar chain level Gal 0 the double-antenna complex N-glycan level Gal 1 with a galactose at the end and the double antenna complex N-glycan level Gal 2 with 2 galactose ends;
  • the method further comprises one or more steps selected from the group consisting of:
  • the product further comprises a kit, device, system, and/or combination thereof for detecting other malignant tumor indicators, preferably the indicator is selected from: a basic condition of the patient, such as age, Gender, family history; clinical manifestations of tumors, including local manifestations such as masses and secondary symptoms, pain, pathological secretions, skin mucosal ulcers, systemic manifestations such as fever, infection, anemia, weight loss, fatigue, dyscrasia, internal diagnosis such as Rectal examination, gynecological examination; specific physical and chemical examination indicators, including specific imaging examinations, such as nuclear magnetic resonance, CT, B-ultrasound, endoscopy, mammography, tumor biochemical indicators, such as AFP, CEA, PSA, SF, TSGF, POA, PROGRP, CA125, CA15-3, CA19-9, CA72-4, CA242, CA50, CYFRA21-1, NSE, SCC, AFU, EBV-VCA, and the like.
  • a basic condition of the patient such as age, Gender,
  • reagents, instruments, modules and/or processors are provided for the preparation of a product for screening, early diagnosis, prognostic evaluation, risk assessment, condition monitoring and/or efficacy evaluation of a subject's malignancy;
  • the product can be, for example, a kit, device, system, and/or combination thereof, the reagent, instrument, module, and/or processor comprising:
  • A Reagents, apparatus, modules and/or processors for determining the level of terminal galactosylation of a blood IgG surface dual antenna complex N sugar chain for the determination of a double antenna complex with no galactose linkage at the IgG surface end Type N-glycan level Gal 0 , double antenna complex N-glycan level Gal 1 with galactose at the end and double antenna complex N-glycan level Gal 2 with 2 galactose at the end;
  • reagents, instruments, modules, and/or processors for one or more methods selected from the group consisting of matrix-assisted laser desorption time-of-flight mass spectrometry MALDI-MS, fast atom bombardment mass spectrometry FAB-MS, electrospray mass spectrometry ESI-MS; liquid chromatography; liquid chromatography-mass spectrometry; sugar chip technology; nuclear magnetic resonance NMR, preferably matrix-assisted laser desorption time-of-flight mass spectrometry MALDI-MS;
  • An optional module or processor for determining whether a subject has a malignancy according to a Gal ratio, a prognostic assessment of the subject, and/or a risk assessment, condition monitoring, and/or efficacy evaluation of the tumor is optionally determining whether a subject has a malignancy according to a Gal ratio, a prognostic assessment of the subject, and/or a risk assessment, condition monitoring, and/or efficacy evaluation of the tumor.
  • the product is as previously described.
  • a method for screening a drug and/or a method of treatment comprising:
  • (A") determining the terminal galactosylation level of an IgG surface dual antenna complex N sugar chain in a blood sample at a different time point before, after, and/or during treatment of the subject, said terminal galactosyl group
  • the levels include: two-antenna complex N-glycan level Gal 0 with no galactose linkage at the end of IgG, two antennas with one galactose at the end, complex N-glycan level Gal 1 and two and a half ends Lactose double antenna complex N - sugar chain level Gal 2 ;
  • (B) calculates the abundance ratio of Gal 0 to Gal 1 and Gal 2 , that is, the Gal ratio;
  • (C) determining the effectiveness of the drug and/or treatment method based on the Gal ratio assessment, wherein the Gal ratio is higher than the Gal ratio obtained in the previous test at or above a predetermined range (eg, 5% to 50%) Or any point or sub-range within the range, indicating that the drug and/or treatment is not effective.
  • a predetermined range eg, 5% to 50%
  • a method for screening, early diagnosis, prognosis, risk assessment, condition monitoring and/or efficacy evaluation of a subject malignant tumor characterized in that the method employs the invention Product or application of the invention.
  • the method comprises:
  • the method further comprises one or more steps selected from the group consisting of:
  • the indicators are selected from: basic conditions of the patient, such as age, sex, family history; tumor clinical Performance, including local manifestations such as mass and secondary symptoms, pain, pathological secretions, skin mucosal ulcers, systemic manifestations such as fever, infection, anemia, weight loss, fatigue, dyscrasia, internal examination such as digital rectal examination, gynecological examination; Specific physical and chemical examination indicators, including specific imaging examinations, such as nuclear magnetic resonance, CT, B-ultrasound, endoscopy, mammography, tumor biochemical indicators, such as AFP, CEA, PSA, SF, TSGF, POA, PROGRP, CA125 , CA15-3, CA19-9, CA72-4, CA242, CA50, CYFRA21-1, NSE, SCC, AFU, EBV-VCA, and the like.
  • basic conditions of the patient such as age, sex, family history
  • tumor clinical Performance including local manifestations such as mass and secondary symptoms, pain, pathological secretions, skin mucosal ulcer
  • the method is for use as described below and includes the corresponding steps as follows:
  • (v) for cancer patient condition monitoring when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a predetermined range (for example, 5% to 50% or any point or sub-range within the range), It indicates that the condition of the cancer patient has developed or worsened;
  • a predetermined range for example, 5% to 50% or any point or sub-range within the range
  • Gal ratios are measured and calculated at different time points before and after treatment or during treatment, when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a preset range (eg 5%) ⁇ 50% or any point or sub-range within the range) indicates that the therapy and/or drug is not effective;
  • the threshold value in (i) to (iv) is 0.35 to 0.6, and more preferably 0.5.
  • the optimal threshold may be determined based on a maximum point of slope in the ROC curve or a value that maximizes sensitivity and specificity, and a preferred threshold range may be an optimal threshold of ⁇ 1 to 15% (including within the range) The range of any numerical point or subrange).
  • the products and applications are used in a method comprising the steps of:
  • the isolated IgG is treated with glycosidase to isolate the N sugar chain of IgG, preferably using glycosidase PNGase F;
  • a mixed solvent of trifluoroacetic acid 25:74.95:0.05 to elute the sugar chain;
  • results of subject malignant tumor screening, early diagnosis, prognosis assessment, risk assessment, condition monitoring and/or efficacy evaluation were obtained based on the target serum IgG surface Gal ratio.
  • the products and/or methods of the invention are individual indicators of subject malignancies screening, early diagnosis, prognostic assessment, risk assessment, condition monitoring, and/or efficacy assessment, ie, not associated with other tumor markers Combined malignant tumor screening, early diagnosis, prognosis assessment, risk assessment, disease monitoring and/or efficacy assessment can be achieved.
  • the products and/or methods of the invention are not combined with the CA125 indicator.
  • the products and/or methods of the invention are not used in the judgment of benign and malignant tumors.
  • the products and/or methods of the invention are not used for the benign and malignant judgment of ovarian diseases, particularly ovarian diseases with elevated CA125 levels.
  • the elevated CA125 level is, for example, >35 U/mL, or > clinically set upper limit of normal.
  • Figure 1 Comparison of various malignant tumor samples with controls (healthy and benign);
  • A representative serum mass IgG N-sugar chain representative mass spectrum (the number above the peak indicates its molecular weight);
  • Figure 2A ROC curves for comparison of samples of various malignant tumors with controls (healthy and benign);
  • Figure 2B ROC curve of ovarian cancer samples compared to controls (healthy and benign);
  • Figure 2C ROC curve of a lung cancer sample compared to a control (healthy person and benign disease);
  • Figure 2D ROC curve of gastric cancer samples compared to controls (healthy humans and benign diseases);
  • Figure 2E ROC curve of liver cancer samples compared to controls (healthy and benign);
  • Figure 2F ROC curve of a comparison of intestinal cancer (including large intestine, small intestine, colon, rectal cancer) samples with controls (healthy humans and benign diseases);
  • Figure 2G ROC curves of comparative samples of various malignant tumors and benign diseases
  • Figure 2H ROC curve of a sample of patients with early malignancy compared to control (healthy and benign disease) samples.
  • Table 1 Control sample classification and sample age distribution
  • Table 2 Staging and grading statistics of malignant tumor samples
  • Table 3A Statistical comparison of Gal ratio detection in malignant tumor patients and control samples
  • Table 3B Comparison of tumor classification and detection rate of various indicators.
  • the present invention is based on the inventors' research in the above-mentioned fields, and provides a method for screening for malignant tumors based on changes in immunoglobulin G (IgG) surface double-antenna complex N-glycan terminal galactosylation in mammalian (preferably human) blood.
  • IgG immunoglobulin G
  • New methods for early diagnosis, prognosis assessment, risk assessment, disease monitoring, and/or efficacy assessment Compared with the existing tumor serological detection method, the method of the invention has wide applicability, high sensitivity and specificity. The characteristics of good sex and convenient operation are of great significance for the early detection of tumors, timely diagnosis and treatment, improvement of prognosis and improvement of survival rate.
  • the purpose of the present invention is to determine the degree of terminal galactosylation of IgG surface double-antenna complex N sugar chains in malignant tumors and control blood (specifically, N sugar chain terminal galactose (Gal 0 ) and galactose chain (Gal 1) According to the difference of Gal 2 ) abundance, the analysis of the degree of galactosylation can distinguish patients with malignant tumors from healthy/benign controls, and achieve early screening, benign and malignant identification, prognosis detection, disease monitoring and / or efficacy evaluation, and thus provide a highly sensitive, highly specific screening method for malignant tumors.
  • the inventors have found that the Gal ratio according to the present invention is generally associated with the occurrence and benign and malignant effects of various tumors.
  • the present invention screens tumors for newly diagnosed patients (liver cancer, stomach cancer, lung cancer, ovarian cancer, intestinal cancer, esophageal cancer, pancreatic cancer, cholangiocarcinoma, renal cancer, breast cancer, prostate cancer). , nasopharyngeal carcinoma, cervical cancer, bladder cancer, endometrial cancer) and differential diagnosis of benign and malignant, with higher sensitivity, specificity and stability; when combined with existing classical indicators, significantly improve the malignant tumor The detection rate.
  • the method of the present invention can be used in combination with classical indicators to increase the detection rate in a particular tumor
  • Gal ratio can be used independently for screening for malignant tumors, early detection of malignant tumors, prognosis of malignant tumors, disease monitoring and/or efficacy evaluation, which can be used alone or in combination with classical indicators for malignant tumors. Check out.
  • containing includes “includes”, “consisting essentially of”, “consisting essentially of”, and “consisting of”; “mainly by...
  • Consisting includes “consisting essentially of” and “consisting of” are subordinate concepts of “contains,” “has,” or “includes.”
  • Gal ratio refers to the amount of terminal galactosylation of a double-antennary complex N-glycan of a subject's blood IgG surface, specifically a two-antenna complex N-glycan end-galactose (Gal 0) a ratio of abundance to galactose chains (Gal 1 and Gal 2 ), where Gal 0 corresponds to a double-antenna complex N-glycan chain with no galactose linkage at the end, and Gal 1 corresponds to a galactose at the end A dual-antenna complex N-glycan chain, Gal 2 corresponds to a two-antenna complex N-glycan chain with two galactose ends.
  • the term "dual antenna complex N-glycan” refers to a branched sugar chain in which two non-pure mannoses are attached to a core pentasaccharide shared in an N-glycan chain, like an antenna (see Cha Xiliang). Chief Editor, Biochemistry, People's Health Publishing House, 7th edition, 2008, pp. 454-455).
  • Gal 0 , Gal 1 and Gal 2 structures are shown below, wherein GlcNAc represents N-acetylglucosamine, Man represents mannose, Gal represents galactose, and Fuc represents fucose.
  • the terms “specificity,” “sensitivity,” and “conformity” have the same meaning as corresponding medical statistical terms. “Specificity” refers to the proportion of cases diagnosed as “non-cancer” that are negative by the kit. “Sensitivity” refers to the proportion of cases that are pathologically diagnosed as “cancer” and which are positive by the kit. “Accuracy” refers to the ratio of the sum of the true positive sample to the true negative sample to the total sample size, reflecting the extent to which the test results of the kit match the true condition of the subject with cancer.
  • threshold As used herein, the terms “threshold”, “cut-off value”, “cutoff value”, “boundary value”, and “reference value” are used interchangeably and refer to the criteria used to determine the test result, ie, the critical value. If the test result is higher than the threshold, it is regarded as positive, and if it is lower than the threshold, it is regarded as negative.
  • CR, PR and SD have exactly the same meanings as the corresponding terms in the solid tumor efficacy evaluation standard (RECIST standard), namely CR (Complete response) tumor complete remission, PR (Partial response) tumor partial remission and SD (Stable disease).
  • RECIST standard solid tumor efficacy evaluation standard
  • the disease is stable.
  • the three indicators CR, PR and SD are defined as clinically beneficial populations, and the opposite is PD (Progressive disease).
  • the term "healthy person/subject” refers to a person/subject having no known benign and malignant diseases, a normal appearance, and no symptoms of any visible disease.
  • malignant tumor patient refers to a patient diagnosed as having a malignant tumor by histopathological reports.
  • the malignant tumor includes but is not limited to: liver cancer, gastric cancer, lung cancer, ovarian cancer, intestinal cancer, esophageal cancer, pancreatic cancer, cholangiocarcinoma, renal cancer, breast cancer, prostate cancer, nasopharyngeal cancer, cervical cancer, bladder cancer, uterus Endometrial cancer.
  • the malignancy is in a stage selected from the group consisting of early cancer, advanced cancer, and the like.
  • non-cancer-related disease patient or “benign disease patient” refers to a patient diagnosed as having no cancer but having a benign related disease by pathological diagnosis (eg, clinical biochemical, imaging, or histopathological report), such as
  • pathological diagnosis eg, clinical biochemical, imaging, or histopathological report
  • patients who are diagnosed as not having lung cancer but suffering from benign lung-related diseases such as pneumonia and tuberculosis are referred to in the liver cancer test as patients who are not diagnosed with liver cancer but have benign liver-related diseases such as hepatitis and cirrhosis.
  • the patient who is diagnosed as a colorectal cancer but has inflammation, colorectal polyps and other benign diseases of the colorectal is considered to be not breast cancer but has benign breast related diseases such as mastitis and breast hyperplasia. patient.
  • non-malignant sample refers to: a healthy person as well as a patient with a benign disease (ie, a patient diagnosed as a cancer by a pathological diagnosis but having a benign related disease).
  • high-risk population/group refers to a population/group with a high degree of risk of developing a tumor, especially a population that is consciously devoid of cancer based on conditions such as age and lifestyle, and is not classified by clinical indicators. in accordance with.
  • the likelihood of developing a tumor in a high-risk population of tumors is much higher than in the general population.
  • lung cancer as an example, males who have smoked for many years are at high risk; in the case of liver cancer, hepatitis B virus infection is a high-risk group.
  • malignant tumor screening means that a healthy population/population is regularly examined by a specific test method, and a potentially healthy person and a subclinical person/subject are identified and expected Through further diagnostic procedures, patients can be detected early, and early treatment can prevent the occurrence of disease or slow down the disability and death caused by the disease, so that patients can obtain a better prognosis and quality of life (Fang Jigan, Liu Qing, "Chinese Oncology", 2002, 11 (1): 10-11).
  • the term "early diagnosis of malignancy” refers to: an early stage patient specifically for the tumor
  • the purpose of the diagnosis and treatment method is to detect early treatment, thereby alleviating the pain, mental and economic burden of the patient, and striving for the early recovery of the tumor patient through early diagnosis and treatment of the tumor.
  • a blood sample of a subject can be collected and separated and preserved whole blood, serum and/or plasma using conventional methods known in the art.
  • the test of the present invention can be carried out using fresh or frozen blood, serum or plasma.
  • the blood sample can be obtained from a variety of mammalian subjects, preferably human, monkey, dog, horse, cow, sheep, pig, mouse, rabbit, and the like. It will be understood by those skilled in the art that due to differences in composition, the Gal ratio may be slightly different when the plasma sample and the serum sample of the subject are separately used for detection. Serum samples are preferred in the present invention for the purpose of facilitating quantification and comparison.
  • IgG isolation kit is commercially available, for example, from Thermo Fisher Scientific.
  • an IgG purification column is used to separate the IgG in the sample, preferably a protein A purification column, and a weak alkaline binding buffer solution and a weak acid elution buffer solution matched with the purification column are used, and more preferably,
  • the purification column is a high throughput purification column, for example, a purification column capable of simultaneously processing 96 samples.
  • N-glycans on IgG surface and their isolation, purification and analysis
  • N sugar chains can be isolated from IgG by methods known in the art including, but not limited to, enzymatic methods, such as the use of glycosidases, preferably glycosidase PNGase F; chemical methods, such as the use of glycoprotein lysis reagents such as ADM0155A ⁇ Solution kit.
  • enzymatic methods such as the use of glycosidases, preferably glycosidase PNGase F
  • chemical methods such as the use of glycoprotein lysis reagents such as ADM0155A ⁇ Solution kit.
  • the N sugar chain can be isolated and/or purified by methods known in the art including, but not limited to, porous graphitized carbon PGC solid phase extraction, polysaccharide purification column , lectin affinity (such as continuous lectin affinity chromatography SLAC), capillary electrophoresis, high performance liquid chromatography.
  • methods known in the art including, but not limited to, porous graphitized carbon PGC solid phase extraction, polysaccharide purification column , lectin affinity (such as continuous lectin affinity chromatography SLAC), capillary electrophoresis, high performance liquid chromatography.
  • PGC porous graphitized carbon
  • the abundance of IgG surface level double antenna complex N sugar chain terminal galactose (Gal 0 ), galactose chain (Gal 1 and Gal 2 ) can be determined by methods known in the art, as long as the method
  • the level of galactosylation in the N-glycan chain can be quantified.
  • the method includes, but is not limited to, mass spectrometry, such as matrix-assisted laser desorption time-of-flight mass spectrometry MALDI-MS, fast atom bombardment mass spectrometry FAB-MS, electrospray ionization mass spectrometry ES-MS; liquid chromatography; liquid chromatography-mass spectrometry Usage; sugar chip technology; nuclear magnetic resonance NMR or any combination of the above.
  • matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) is preferably employed for high-throughput rapid detection of the degree of terminal galactosylation of IgG surface dual-antenna complex N-glycans.
  • the derivatization step of the sugar chain is not required, which not only simplifies the analysis step, but also saves the analysis time, especially avoiding the experimental error caused by the derivatization step;
  • matrix-assisted laser desorption time-of-flight mass spectrometry MALDI-MS is employed, preferably the quality calibration standard used in the test is TOFmix, the substrate used is 2,5-dihydroxybenzoic acid DHB and/or used
  • the homogenization reagent is ethanol.
  • the Gal ratio that is, the abundance ratio of the terminal galactose (Gal 0 ) to the galactose chain (Gal 1 and Gal 2 ) of the double-antennary complex N sugar chain of the subject blood IgG surface, as a malignant tumor Indicators for screening, early diagnosis, prognostic assessment, risk assessment, disease monitoring, and/or efficacy assessment.
  • the abundance ratio of Gal 0 to Gal 1 and Gal 2 can be calculated using the following formula:
  • Gal ratio aGal 0 / (bGal 1 + cGal 2 ), or other reciprocal, logarithmic and other transformations.
  • a, b, and c are independently between 0 and 10, and a ⁇ 0, b and c are not 0 at the same time.
  • a, b, and c are each independently selected from the group consisting of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.2, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9 or 10.
  • the specific values of a, b or c can be adjusted according to the different response levels of the test methods and instruments used by Gal 0 , Gal 1 and Gal 2 .
  • the specific values of a, b or c can be adjusted based on the ratio of the test method used to the standard method after verification.
  • a standard method commonly used in the art is Ultra High Performance Liquid Chromatography (UPLC), which can be normalized or weighted by other methods as measured by the standard method.
  • UPLC Ultra High Performance Liquid Chromatography
  • Gal 0 , Gal 1 and Gal 2 Since the total amount of Gal 0 , Gal 1 and Gal 2 itself is not fixed, and Gal 0 , Gal 1 and Gal 2 have some inconsistencies in response to different detection instruments and/or methods, it may be necessary in practical applications.
  • the weighting parameters of Gal 0 , Gal 1 and Gal 2 ie a, b and c) are corrected to ensure that the results meet the detection requirements (eg with the required sensitivity, accuracy, etc.).
  • the formula can be selected from:
  • Gal 0 , Gal 1 and Gal 2 for the abundance ratio are preferably derived from the same sample, the same detection and/or the same spectrum. Therefore, the use of this index can avoid deviations caused by the operation of parallel samples, thus reducing the error of parallel samples entering mass spectrometry, ensuring high reproducibility and accuracy of the analysis, ensuring that the method can be applied to clinical applications. Tumor screening, prognosis assessment, risk assessment, condition monitoring and/or efficacy assessment.
  • a person skilled in the art can draw a corresponding ROC curve according to the detection data of different object groups based on common knowledge in the art.
  • the corresponding ROC curve is drawn according to data of different populations.
  • Each ROC curve provides a series of thresholds and corresponding sensitivity and specificity, according to techniques well known in the art. Therefore, using these ROC curves, those skilled in the art can easily find out the ability of the detection to identify diseases when selecting any threshold (ie, cut-off value), that is, to diagnose the sensitivity, specificity, and coincidence rate of the corresponding cancer species. .
  • the calculation of the screening results depends on the specificity and sensitivity.
  • the calculation method of the threshold includes but is not limited to:
  • Method 1 Select the maximum slope of the ROC curve and take the corresponding Gal ratio as the threshold.
  • Method 2 Maximize the value of sensitivity and specificity, that is, the Gal ratio corresponding to [sensitivity%-(1-specificity%)] max is the optimal threshold (the most approximate index, Youden index).
  • a preferred threshold range in the present invention is from 0.35 to 0.6, preferably 0.5.
  • the selection of the threshold and the corresponding sensitivity, specificity and coincidence rate parameters may be different, but they are all included in the threshold range defined by the present invention.
  • One skilled in the art can select an appropriate preferred threshold from within the ROC curve or threshold range disclosed herein, based on actual usage requirements, such as requirements for a particular tumor species, sensitivity, specificity, and the like.
  • the optimal threshold may be within the range of ⁇ 1 to 15% (including any numerical or sub-range within the range) after determining the optimal threshold.
  • the Gal ratio indicator of the present invention can be applied to malignant tumor screening, early diagnosis, prognosis evaluation, risk assessment, disease monitoring, and/or therapeutic evaluation. These applications include but are not limited to:
  • Gal ratio indicators for high-risk group malignant tumors is expected: when the Gal ratio reaches or exceeds a predetermined threshold, the probability of a malignant tumor is higher than other subjects;
  • the Gal ratio indicator is used for cancer patient condition monitoring: when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a preset range (for example, 5% to 50% or any point or sub-range within the range) Scope), indicating that the condition of the cancer patient has developed or further deteriorated;
  • a preset range for example, 5% to 50% or any point or sub-range within the range
  • the Gal ratio indicator is used for the evaluation of the efficacy: the Gal ratio is measured and calculated at different time points before and after treatment or during the treatment, when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds the preset range ( For example, 5% to 50% or any point or sub-range within the range indicates poor efficacy of the therapy and/or drug.
  • Methods and products for screening for malignant tumors, early diagnosis, prognostic assessment, risk assessment, condition monitoring, and/or efficacy evaluation are provided in the present invention based on the techniques described above. .
  • the method of the invention comprises the following steps:
  • the sugar chain level Gal 0 the double-antenna complex N-glycan level Gal 1 with a galactose at the end and the double antenna complex N-glycan level Gal 2 with 2 galactose ends;
  • Step (C') may include, in different applications:
  • Gal ratio For tumor screening detection: When the Gal ratio reaches or exceeds a predetermined threshold, the subject is determined to have a malignancy or is at risk of developing a malignant tumor.
  • Gal ratio reaches or exceeds a predetermined threshold, it is determined that the subject has a malignant tumor or is at risk of suffering from a malignant tumor.
  • the target group (such as high-risk group) malignant tumor occurrence is expected: when the Gal ratio reaches or exceeds a preset threshold, the probability of presenting a malignant tumor is higher than other subjects;
  • the Gal ratio is measured and calculated at different time points before and after treatment or during treatment, when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds the preset range (for example, 5% ⁇ 50% or any point or sub-range within the range) indicates that the therapy and/or drug is not effective.
  • the preset range for example, 5% ⁇ 50% or any point or sub-range within the range
  • the threshold in the method of the invention may range from 0.35 to 0.6, preferably 0.5.
  • the threshold range can be adjusted depending on the subject population and the tumor type, such as determining the corresponding threshold range based on the maximum point of the slope in the ROC curve or the value of maximizing sensitivity and specificity.
  • the method of the invention may optionally comprise one or more steps selected from the group consisting of:
  • step (h') can be performed as follows:
  • (i') for tumor screening detection when the Gal ratio reaches or exceeds a predetermined threshold, the subject is determined to have a malignancy or is at risk of developing a malignant tumor;
  • (iii') for prognostic monitoring when the Gal ratio reaches or exceeds a predetermined threshold, it indicates that the subject has a high risk of recurrence or recurrence;
  • (v') for cancer patient condition monitoring when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a preset range (for example, 5% to 50% or any point or sub-range within the range) , indicating that the condition of the cancer patient has developed or worsened;
  • a preset range for example, 5% to 50% or any point or sub-range within the range
  • the Gal ratio is measured and calculated at different time points before and after treatment or during treatment, when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a preset range (for example, 5 % to 50% or any point or sub-range within the range) indicates that the therapy and/or drug is not effective.
  • a preset range for example, 5 % to 50% or any point or sub-range within the range
  • the threshold value in (i') to (iv') may be 0.35 to 0.6, preferably 0.5.
  • the threshold can also be set by a person of ordinary skill in the art according to specific needs.
  • the method of the invention is preferably a high throughput detection method, for example, 96, 192, 288, 384, 480, 576 samples can be processed simultaneously.
  • the present invention also provides a product for screening for malignant tumors, early diagnosis, prognosis evaluation, risk assessment, condition monitoring, and/or efficacy evaluation.
  • the product can be, for example, a kit, device, system, and/or combination thereof.
  • the products include:
  • (C) optional, for judging whether the subject has a malignant tumor according to the Gal ratio, and prognosing the subject
  • product of the present invention optionally includes one or more selected from the group consisting of:
  • (v) for cancer patient condition monitoring when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a predetermined range (for example, 5% to 50% or any point or sub-range within the range), It indicates that the condition of the cancer patient has developed or worsened;
  • a predetermined range for example, 5% to 50% or any point or sub-range within the range
  • Gal ratios are measured and calculated at different time points before and after treatment or during treatment, when the Gal ratio is higher than the Gal ratio obtained in the previous test reaches or exceeds a preset range (eg 5%) ⁇ 50% or any point or sub-range within the range) indicates that the therapy and/or drug is not effective.
  • a preset range eg 5%
  • the threshold value in (i) to (iv) may be 0.35 to 0.6, preferably 0.5.
  • the threshold can also be set by a person of ordinary skill in the art according to specific needs.
  • the invention meets the operational requirements that the excellent clinical diagnostic index should meet in the actual detection process, and shows that the blood ratio of detecting blood IgG has practical feasibility in clinical application of malignant tumor screening, early diagnosis, prognosis evaluation and risk assessment.
  • malignant tumors including liver cancer, gastric cancer, lung cancer, ovarian cancer, colon cancer, esophageal cancer, pancreatic cancer, cholangiocarcinoma, kidney cancer, breast cancer, prostate cancer, nasopharyngeal cancer, cervical cancer, bladder) Cancer, Endometrial cancer;
  • the present invention has high specificity and sensitivity in malignant tumor screening, thereby judging that the methods and indicators of the present invention have clinical significance.
  • Blood samples from healthy people, benign control blood samples, preoperative blood samples from malignant tumors, blood samples from patients with precancerous lesions, and postoperative blood samples from malignant tumors were taken from hospitalized and outpatients in tertiary hospitals.
  • Malignant tumors and precancerous lesions serum samples with clear pathological diagnosis of tumors or patients with clear precancerous lesions; among them, the staging and grading statistics of tumor samples are shown in Table 2, and the classification statistics are shown in Table 3. (referred to in the "Malignant Tumor INM Classification" manual).
  • the TNM staging standard is commonly used internationally.
  • the TNM classification method is based on clinical findings for the pathological staging of cancer:
  • T the size of the primary tumor (initial cancer), the extent of infiltration, the presence or absence of metastasis, and the depth of infiltration, are divided into five levels (T0 to T4). The higher the number, the more obvious the cancer progresses. The classification methods based on the different organs in which cancer occurs are also different.
  • N the degree of metastasis of the surrounding lymph nodes, divided into four levels (N0 to N3), the greater the number, the more obvious the progression of cancer.
  • M0 means that no other organ transfer occurred
  • M1 means that other organ transfer has occurred
  • Stage I The tumor is confined to one place with no signs of spreading.
  • Stage II The tumor has spread to adjacent lymph nodes but does not affect other organs or tissues.
  • Stage III In addition to spreading to adjacent lymph nodes, tumors also affect nearby organs or tissues.
  • Stage IV The tumor has spread to distant parts. The earlier the period, the better the treatment effect, and the longer the patient survives. If surgical methods are used to remove cancer, the excised cancer tissue specimens should be examined in detail after surgery, and the TNM staging should be performed after surgery.
  • Pathological grading of cancer can be based on the degree of differentiation of cancer cells:
  • Grade I is well differentiated and has a low degree of malignancy
  • Grade II is moderately differentiated and moderately malignant
  • Grade III is poorly differentiated and has a high degree of malignancy.
  • the pathological grade of malignant tumors suggests the degree of malignancy of the tumor.
  • Samples of healthy people People without any known benign and malignant diseases, with normal appearance and without any visible disease symptoms.
  • Benign control samples A sample of patients diagnosed with benign disease rather than malignant tumors by clinical biochemical, imaging, or histopathological reports.
  • Malignant tumor patient samples A sample of patients diagnosed with malignant tumors by histopathological reports.
  • High-risk population sample A sample of a population at high risk of developing a tumor. The likelihood of developing a tumor in a high-risk population of tumors is much higher than in the general population. Taking lung cancer as an example, males who have smoked for many years are at high risk; in the case of liver cancer, hepatitis B virus infection is a high-risk group.
  • Sample preservation 5 mL of venous blood was drawn, and after coagulation (still standing in a coagulation tube at room temperature for 30 minutes), centrifuged at 2000 g for 10 minutes, the upper serum was aspirated, and stored at -80 °C for later use.
  • Serum detection and data processing were performed under double-blind conditions.
  • Protein A Spin Plate for IgG Screening (96 well) was purchased from Thermo Fisher Scientific, Cat. No. 45202.
  • the binding buffer solution, the elution buffer solution, and the BCA kit were purchased from Thermo Fisher Scientific.
  • Glycosidase PNGase F (500 U/ ⁇ L) was purchased from New England Biolabs
  • Porous graphitized carbon PGC purchased from Grace Corporation.
  • Mass spectrometer Axima Resonance MALDI-QIT-TOF mass spectrometer (Shimadzu Corporation).
  • Homogeneous reagent ethanol, HPLC grade (Merck).
  • Matrix 2,5-dihydroxybenzoic acid DHB, purchased from Sigma-Aldrich.
  • the purification column and the buffer solution (binding buffer solution and rinse buffer solution) are equilibrated at room temperature for 30 minutes;
  • PLC Porous graphitized carbon
  • a 96-well plate containing graphitized carbon is activated by an 80% aqueous solution of acetonitrile containing 0.1% trifluoroacetic acid;
  • the sugar chain was eluted with a 25% aqueous solution of acetonitrile containing 0.05% trifluoroacetic acid and the eluted solution was collected for analysis.
  • MALDI mass spectrometry conditions positive ion reflection mode; 337 nm nitrogen laser source; laser energy set to 125 V; sample acquisition mode is “auto experiment”; each MS spectrum is cumulative 200 profiles, 2 shot/profile; acquisition m/ The z range is 500-3000.
  • the method for judging the degree of galactosylation was used to quantitatively analyze the degree of galactose at the end of the complex IgG chain of the IgG surface of the IgG surface of the screening target, and calculate the double antenna complexity of the serum according to the peak height or the size of the peak area.
  • the ratio of the galactose to galactose chain abundance (Gal ratio, Gal ratio) at the end of the N-type sugar chain, and used as an indicator of the galactose content of the IgG surface.
  • the calculation formula used in the following examples is:
  • Gal ratio [Gal 0 / (Gal 1 + 2 ⁇ Gal 2 )]
  • ROC curve is commonly used in clinical practice to compare and determine the ability of a diagnostic indicator to recognize a disease.
  • AUC area-under-the-curve
  • the diagnostic indicator is considered to be “highly accurate”; when the AUC is between 0.7-0.9 and 0.5-0.7, it is “applicable” and “invalid respectively”.
  • a threshold of 0.5 (the judgments made in other examples below are also based on the threshold) is used as a criterion for determining whether the subject has a malignant tumor or has a risk of developing a malignant tumor, and as a result (see Table 3B):
  • the detection rate of malignant tumors can reach about 88.72%
  • liver cancer cases Gal ratio and AFP were used as independent diagnostic indicators, the detection rates were 93.98% and 59.72%, respectively. 87 cases of liver cancer serum samples with AFP negative ( ⁇ 20ng/ml) were detected, and the method of the present invention can detect 72 cases;
  • Gal ratio and CA125 were used as independent diagnostic indicators, the detection rates were 92.34% and 69.79%, respectively.
  • the method of the present invention 218 cases can be detected.
  • cancers include breast cancer, nasopharyngeal cancer, cervical cancer, bladder cancer, and endometrial cancer.
  • the joint indicator means that the Gal ratio indicator of the present invention is combined with the existing classical index for testing
  • the Gal method is highly sensitive and highly specific and can be effectively utilized for screening of malignant tumors, and the method and index of the present invention have higher accuracy than the existing classical standards. Moreover, the combination of the methods and indicators of the present invention with existing classical standards can further increase the detection rate of malignant tumors.
  • Example 2 Gal ratio is used to distinguish between ovarian cancer patients and controls (healthy and benign diseases)
  • the above analysis confirmed that the serum IgG galactosylation index of the present invention can effectively screen and distinguish ovarian cancer patients and controls with high sensitivity and specificity.
  • the mean Gal ratio of lung cancer patients was significantly different from the control sample (t test, p ⁇ 0.0001) (see Table 3A).
  • the mean Gal ratio of liver cancer patients was significantly different from the control sample (t test, p ⁇ 0.0001) (see Table 3A).
  • the above analysis confirmed that the serum IgG galactosylation index of the present invention can effectively screen and distinguish liver cancer and control with high sensitivity and specificity.
  • the above analysis confirmed that the serum IgG galactosylation index of the present invention can effectively screen and distinguish intestinal cancer and control with high sensitivity and specificity.
  • Example 7 Gal ratio is used to distinguish various types of malignant tumors from benign controls
  • Example 8 Gal has a higher diagnostic value than combined with classical indicators
  • the detection rate in a specific tumor can be improved.
  • Gal was used in combination with AFP, and the detection rate reached 95.83% (one of the two indicators was positive), which was higher than the Gal ratio (93.98%) and AFP (59.72%).
  • Detection In 158 patients with gastric cancer, Gal was used in combination with CEA, and the detection rate reached 91.14%, which was higher than that of Gal ratio (88.61%) and CEA (59.49%).
  • Gal ratio can be used in combination with classical indicators to increase the detection rate of malignant tumors.
  • a follow-up monitoring study was performed on 78 patients with gastric cancer who underwent standardized surgical procedures to examine the correlation between the Gal ratio in the blood and the patient's condition. The detection period was once every 3 months.
  • Postoperative tumor recurrence was evaluated based on pathological diagnosis, clinical biochemistry, and imaging data. The results showed that there were 25 recurrences within 3 years after surgery, and 23 of them were positive for Gal before the diagnosis of tumor recurrence (detection rate was 92.0%).
  • Example 11 Gal ratio is used to predict the occurrence of malignant tumors
  • Gal ratio detection can be used to predict the occurrence of malignant tumors.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Biophysics (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Optics & Photonics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un produit utilisé pour permettre un criblage des tumeurs malignes, un diagnostic précoce, une évaluation prognostique, une évaluation des risques, une surveillance de l'état et/ou une évaluation de l'efficacité, une application d'un réactif, un instrument, un module et/ou un appareil de traitement dans la préparation du produit, ainsi qu'un procédé utilisé pour un criblage des médicaments et/ou d'un procédé de traitement. Le produit comprend : (A) un réactif, un instrument et/ou un système utilisés pour déterminer le degré de galactosylation terminale de chaîne de N-sucre de complexe bi-antennaire à la surface des IgG dans le sang, déterminer une abondance de chaîne de N-sucre de complexe bi-antennaire sans liaison de galactose terminal à la surface des IgG (Gal0), une abondance de chaîne de N-sucre de complexe bi-antennaire à simple liaison de galactose terminal (Gal1) et une abondance de chaîne de N-sucre de complexe bi-antennaire à double liaison de galactose terminal (Gal2); (B) un module et/ou un dispositif de traitement utilisés pour calculer une valeur de rapport d'abondance de Gal0, de Gal1 et de Gal2 (rapport des Gal); (C) facultativement, un module et/ou un dispositif de traitement utilisés pour déterminer si une cible est atteinte d'une tumeur maligne et effectuer une évaluation prognostique et/ou une évaluation du risque de tumeurs, une surveillance de l'état et/ou une évaluation de l'efficacité sur la cible en fonction du rapport des Gal.
PCT/CN2016/100048 2015-09-29 2016-09-26 Produit, application et procédé utilisés pour permettre une évaluation et un criblage se rapportant à des tumeurs malignes WO2017054686A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201510632081.1A CN105277718B (zh) 2015-09-29 2015-09-29 用于恶性肿瘤相关筛查及评估的产品、应用及方法
CN201510632081.1 2015-09-29

Publications (1)

Publication Number Publication Date
WO2017054686A1 true WO2017054686A1 (fr) 2017-04-06

Family

ID=55147086

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/100048 WO2017054686A1 (fr) 2015-09-29 2016-09-26 Produit, application et procédé utilisés pour permettre une évaluation et un criblage se rapportant à des tumeurs malignes

Country Status (2)

Country Link
CN (1) CN105277718B (fr)
WO (1) WO2017054686A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110927263A (zh) * 2019-10-21 2020-03-27 复旦大学附属妇产科医院 利用血清糖基化修饰联合ca125预测卵巢癌患者铂类化疗敏感性的方法
CN111781357A (zh) * 2020-07-10 2020-10-16 万东山 一种同时检测肺癌四种肿瘤标志物的试剂盒与应用
CN113808741A (zh) * 2021-10-08 2021-12-17 国家癌症中心 一种癌症防控智能网络平台
CN117131468A (zh) * 2023-09-12 2023-11-28 中山大学孙逸仙纪念医院 肝内胆管细胞癌筛查指标及其预后因素的分析评价方法

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105277718B (zh) * 2015-09-29 2018-03-20 上海知先生物科技有限公司 用于恶性肿瘤相关筛查及评估的产品、应用及方法
CN105759062B (zh) * 2016-03-29 2018-03-27 复旦大学附属中山医院 一种基于生物标志物谱针对中国农村人口高危人群的肺癌风险预测试剂盒
CN106404975B (zh) * 2016-09-22 2018-07-17 谱天（天津）生物科技有限公司 一种个体化药物的筛选方法及其应用
CN108334887B (zh) * 2017-01-19 2023-06-13 腾讯科技（深圳）有限公司 一种用户选取方法和装置
CN106596827B (zh) * 2017-01-23 2019-06-07 上海知先生物科技有限公司 Gal指标在自身免疫疾病治疗敏感性和疗效评估中的应用
CN107478712A (zh) * 2017-08-07 2017-12-15 章晓联 一种血清IgG的Gal指标在结核病诊断中的应用
CN107817353B (zh) * 2017-11-01 2020-10-13 复旦大学 预测肿瘤坏死因子抑制剂疗效的试剂和方法
CN108504555B (zh) * 2018-04-16 2021-08-03 图灵人工智能研究院（南京）有限公司 鉴别及评价肿瘤进展的装置和方法
CN108982856B (zh) * 2018-07-18 2021-04-02 深圳格道糖生物技术有限公司 基于唾液特异糖蛋白糖链结构的肝癌相关筛查/评估的产品及应用
CN109767844A (zh) * 2018-12-28 2019-05-17 郑州大学第一附属医院 靶向肿瘤介入治疗剂及其制备方法和智能检测系统
CN111366633A (zh) * 2020-01-19 2020-07-03 深圳格道糖生物技术有限公司 基于唾液特异糖蛋白糖链结构的肺部良性疾病筛查、评估的产品及应用
CN112309571B (zh) * 2020-10-30 2022-04-15 电子科技大学 数字病理图像的预后量化特征的筛选方法
CN112450982B (zh) * 2020-12-09 2023-03-28 上海市闵行区中心医院 一种通过b超表现预测前列腺癌的定性和数据评分系统
CN113702482B (zh) * 2021-08-30 2022-08-23 中国医学科学院北京协和医院 一种IgG N-糖链特征组合及其应用
CN115184619B (zh) * 2022-08-01 2023-02-03 中国医学科学院北京协和医院 子宫内膜癌术前筛查及诊断的标志物及其应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040191256A1 (en) * 1997-06-24 2004-09-30 Genentech, Inc. Methods and compositions for galactosylated glycoproteins
CN101074951A (zh) * 2007-06-26 2007-11-21 大连医科大学 一种早期诊断类风湿关节炎的免疫球蛋白g糖基化检测试剂及制备方法
WO2009013538A2 (fr) * 2007-07-20 2009-01-29 National Institute For Bioprocessing Research And Training Marqueurs de glycosylation pour le cancer et l'inflammation chronique
CN105277718A (zh) * 2015-09-29 2016-01-27 上海知先生物科技有限公司 用于恶性肿瘤相关筛查及评估的产品、应用及方法

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020045207A1 (en) * 1997-10-31 2002-04-18 Lynne A. Krummen Glycoprotein production process
WO2006114661A1 (fr) * 2005-04-26 2006-11-02 Dwek Raymond A Analyse de glycane a haut rendement pour diagnostiquer et surveiller l'arthrite rhumatoide et d'autres maladies auto-immunes
US20110117601A1 (en) * 2007-08-31 2011-05-19 Markus Haberger Glycosylation Profile Analysis
CN102250852B (zh) * 2011-06-11 2013-06-12 内蒙古医学院 牛重组α-1,3-半乳糖苷转移酶的克隆与表达
WO2013021279A2 (fr) * 2011-08-10 2013-02-14 Lfb Biotechnologies Anticorps fortement galactosylés
CA2862925C (fr) * 2012-02-10 2020-01-21 University Of Maryland, Baltimore Glyco-ingenierie chimio-enzymatique d'anticorps et de leurs fragments fc

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040191256A1 (en) * 1997-06-24 2004-09-30 Genentech, Inc. Methods and compositions for galactosylated glycoproteins
CN101074951A (zh) * 2007-06-26 2007-11-21 大连医科大学 一种早期诊断类风湿关节炎的免疫球蛋白g糖基化检测试剂及制备方法
WO2009013538A2 (fr) * 2007-07-20 2009-01-29 National Institute For Bioprocessing Research And Training Marqueurs de glycosylation pour le cancer et l'inflammation chronique
CN105277718A (zh) * 2015-09-29 2016-01-27 上海知先生物科技有限公司 用于恶性肿瘤相关筛查及评估的产品、应用及方法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KRISTEL, K. ET AL.: "Immunoglobulin G Fc N-Glycan Profiling in Patients with Gastric Cancer by LC-ESI-MS: Relation to Tumor Progression and Survival", GLYCOCONJUGATE JOURNAL, vol. 29, no. 1, 17 December 2011 (2011-12-17), pages 57 - 66, XP035002293 *
QIAN, YIFAN ET AL.: "Quantitative Analysis of Serum IgG Galactosylation Assists Differential Diagnosis of Ovarian Cancer", JOURNAL OF PROTEOME RESEARCH, vol. 12, no. 9, 6 September 2013 (2013-09-06), pages 4046 - 4055 *
RADKA, S. ET AL.: "Ovarian Cancer is Associated with Changes in Glycosylation in both Acute-Phase Proteins and IgG", GLYCOBIOLOGY, vol. 17, no. 12, 20 September 2007 (2007-09-20), pages 1344 - 1356, XP055372415 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110927263A (zh) * 2019-10-21 2020-03-27 复旦大学附属妇产科医院 利用血清糖基化修饰联合ca125预测卵巢癌患者铂类化疗敏感性的方法
CN111781357A (zh) * 2020-07-10 2020-10-16 万东山 一种同时检测肺癌四种肿瘤标志物的试剂盒与应用
CN111781357B (zh) * 2020-07-10 2024-01-19 万东山 一种同时检测肺癌四种肿瘤标志物的试剂盒与应用
CN113808741A (zh) * 2021-10-08 2021-12-17 国家癌症中心 一种癌症防控智能网络平台
CN113808741B (zh) * 2021-10-08 2022-04-22 国家癌症中心 一种癌症防控智能网络平台
CN117131468A (zh) * 2023-09-12 2023-11-28 中山大学孙逸仙纪念医院 肝内胆管细胞癌筛查指标及其预后因素的分析评价方法
CN117131468B (zh) * 2023-09-12 2024-01-23 中山大学孙逸仙纪念医院 肝内胆管细胞癌筛查指标及其预后因素的分析评价方法

Also Published As

Publication number Publication date
CN105277718B (zh) 2018-03-20
CN105277718A (zh) 2016-01-27

Similar Documents

Publication Publication Date Title
WO2017054686A1 (fr) Produit, application et procédé utilisés pour permettre une évaluation et un criblage se rapportant à des tumeurs malignes
Park et al. Metabolomic-derived novel cyst fluid biomarkers for pancreatic cysts: glucose and kynurenine
Poté et al. Imaging mass spectrometry reveals modified forms of histone H4 as new biomarkers of microvascular invasion in hepatocellular carcinomas
Lokhov et al. Metabolite profiling of blood plasma of patients with prostate cancer
WO2023082821A1 (fr) Marqueur métabolique sérique pour le diagnostic des nodules pulmonaires bénins et malins et son utilisation
Starodubtseva et al. Label‐free cervicovaginal fluid proteome profiling reflects the cervix neoplastic transformation
EP2525227A1 (fr) Procédé de détection du cancer du pancréas en utilisant le marquer sérologique ULBP2
CA2700125A1 (fr) Detection dans la salive, de proteines modulees produites suite a un carcinome canalaire in situ du sein
US8455624B2 (en) Methods and compositions for detecting endometrial or ovarian cancer
Fan et al. Tubulin beta chain, filamin A alpha isoform 1, and cytochrome b-c1 complex subunit 1 as serological diagnostic biomarkers of esophageal squamous cell carcinoma: a proteomics study
Liu et al. Potential biomarkers for esophageal carcinoma detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
Wu et al. Identification of serum biomarkers for ovarian cancer using MALDI–TOF-MS combined with magnetic beads
CN116879558B (zh) 卵巢癌诊断标志物、检测试剂及检测试剂盒
US20130102011A1 (en) Human leucine-rich a-2-glycoprotein-1 and aminopeptidase n as risk indicators for cancer
KR20160146102A (ko) N-당쇄 질량분석을 이용한 대장암 진단방법
CA2705044A1 (fr) Procedes pour detecter ou surveiller un cancer a l'aide de lpc en tant que marqueur
JP5867834B2 (ja) 肺癌マーカー補体Ｃ３ｄｇ分子及び肺癌マーカーの分析方法
WO2021045180A1 (fr) Marqueur de cancer gastrique et procédé d'examen l'utilisant
CN115078719A (zh) 用于结肠癌筛查和诊断的IgG糖链标志物及其应用
WO2020013097A1 (fr) Chaîne de sucre spécifique du cancer de la prostate et procédé de test l'utilisant
Wilz et al. Development of a test to identify bladder cancer in the urine of patients using mass spectroscopy and subcellular localization of the detected proteins
WO2024048514A1 (fr) Méthode de dépistage du cancer du col de l'utérus et/ou de la néoplasie cervicale intra-épithéliale
CN117589991B (zh) 一种用于乳腺癌患者her2表达状态鉴定的生物标志物、模型、试剂盒及用途
KR20180117918A (ko) Mfap5 측정 제제를 포함하는 대장암 진단용 조성물, 키트, 및 이를 이용한 대장암 진단 방법
US20150044703A1 (en) Methods and compositions for detecting endometrial or ovarian cancer

Legal Events

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

Ref document number: 16850310

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 23/07/2018)

122 Ep: pct application non-entry in european phase

Ref document number: 16850310

Country of ref document: EP

Kind code of ref document: A1