CN113189335A - 血清淀粉样蛋白a2在制备检测肺癌试剂盒中的应用 - Google Patents
血清淀粉样蛋白a2在制备检测肺癌试剂盒中的应用 Download PDFInfo
- Publication number
- CN113189335A CN113189335A CN202110671159.6A CN202110671159A CN113189335A CN 113189335 A CN113189335 A CN 113189335A CN 202110671159 A CN202110671159 A CN 202110671159A CN 113189335 A CN113189335 A CN 113189335A
- Authority
- CN
- China
- Prior art keywords
- lung cancer
- serum amyloid
- kit
- saa2
- detecting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102100032007 Serum amyloid A-2 protein Human genes 0.000 title claims abstract description 66
- 206010058467 Lung neoplasm malignant Diseases 0.000 title claims abstract description 53
- 201000005202 lung cancer Diseases 0.000 title claims abstract description 53
- 208000020816 lung neoplasm Diseases 0.000 title claims abstract description 53
- 101000637821 Homo sapiens Serum amyloid A-2 protein Proteins 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 238000001514 detection method Methods 0.000 title abstract description 19
- 238000012216 screening Methods 0.000 claims abstract description 22
- 230000014509 gene expression Effects 0.000 claims abstract description 20
- 239000003147 molecular marker Substances 0.000 claims abstract description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims description 24
- 108010001336 Horseradish Peroxidase Proteins 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 8
- 238000008157 ELISA kit Methods 0.000 claims description 7
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 6
- 239000003085 diluting agent Substances 0.000 claims description 6
- 210000005259 peripheral blood Anatomy 0.000 claims description 4
- 239000011886 peripheral blood Substances 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 239000012089 stop solution Substances 0.000 claims description 2
- 101710083332 Serum amyloid A-2 protein Proteins 0.000 abstract description 39
- 208000002154 non-small cell lung carcinoma Diseases 0.000 abstract description 11
- 238000003745 diagnosis Methods 0.000 abstract description 8
- 101150089320 SAA2 gene Proteins 0.000 abstract description 4
- 239000003814 drug Substances 0.000 abstract description 2
- 239000003550 marker Substances 0.000 abstract description 2
- 238000012795 verification Methods 0.000 abstract description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 15
- 239000002953 phosphate buffered saline Substances 0.000 description 15
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 238000002965 ELISA Methods 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 201000005249 lung adenocarcinoma Diseases 0.000 description 6
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 6
- 206010003445 Ascites Diseases 0.000 description 5
- 238000007865 diluting Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 229920001213 Polysorbate 20 Polymers 0.000 description 4
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 4
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 4
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000003118 sandwich ELISA Methods 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- 206010009900 Colitis ulcerative Diseases 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 206010040047 Sepsis Diseases 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 201000006704 Ulcerative Colitis Diseases 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 208000037815 bloodstream infection Diseases 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000013211 curve analysis Methods 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000439 tumor marker Substances 0.000 description 2
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 239000005635 Caprylic acid (CAS 124-07-2) Substances 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 238000012286 ELISA Assay Methods 0.000 description 1
- 206010041067 Small cell lung cancer Diseases 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000011091 antibody purification Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 208000003849 large cell carcinoma Diseases 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229960002446 octanoic acid Drugs 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 208000000587 small cell lung carcinoma Diseases 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
-
- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/581—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)
-
- 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
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/775—Apolipopeptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种血清淀粉样蛋白A2在制备检测肺癌试剂盒中的应用，属于生物技术和医学领域。本发明具体公开了一种血清淀粉样蛋白A2作为分子标志物在制备检测肺癌试剂盒或试剂中的应用，所述血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。本发明通过试验验证，首次揭示了SAA2基因高表达与肺癌临床分期密切关联，发现SAA2可作为检测NSCLC患者组织和血浆样本的标志物，并开发了可用于临床的肺癌早期筛查和临床分期的快速检测试剂或试剂盒，从而为肺癌诊断提供新的手段。
Description
技术领域
本发明涉及生物技术和医学领域，特别是涉及一种血清淀粉样蛋白A2在制备检测肺癌试剂盒中的应用。
背景技术
肺癌是目前最常见的恶性肿瘤之一，也是全球发病率和死亡率增加最快的肿瘤，每年大概有120万患者被诊断为肺癌，110万人死于肺癌。肺癌依据其组织病理学可分为两大类：小细胞肺癌和非小细胞肺癌。非小细胞肺癌又包含鳞状细胞癌、腺癌、大细胞癌等。
迄今为止，仍然缺乏有效的肺癌早期筛查和临床分期的分子检测手段。开发新的早期筛查和诊断技术己经成为肺癌防治急需解决的重大科学问题。肿瘤标志物的发现和合理应用是肿瘤早期发现、早期诊断的前提。
基于外周血中蛋白标记物可以发展为早期癌症诊断的替代手段。SAA2蛋白在人原代单核细胞和巨噬细胞中表达，并在促进Th17介导的炎症性疾病中发挥独特作用。已发现SAA2可有效区分感染状态和正常状态，并有可能成为评估细菌血流感染(BSI)发生和发展的生物标志物。SAA2可用作诊断或治疗溃疡性结肠炎(UC)和肝细胞癌(HCC)的潜在标志物。
目前SAA2对于肺癌的筛查和临床分期还没有商业化试剂盒。因此本发明开发出可供临床应用的检测试剂，以深入地研究和大规模验证SAA2与肺癌的相关性，对癌症进行诊断及为个体患者提供更好的辅助治疗方法。
发明内容
本发明的目的是提供一种血清淀粉样蛋白A2(SAA2)在制备检测肺癌试剂盒中的应用，以解决上述现有技术存在的问题，首次揭示了SAA2基因高表达与肺癌临床分期密切关联，并开发了可用于临床的肺癌早期筛查和临床分期的快速检测试剂或试剂盒，从而为肺癌诊断提供新的手段。
为实现上述目的，本发明提供了如下方案：
本发明提供一种血清淀粉样蛋白A2作为分子标志物在制备检测肺癌试剂盒中的应用,所述血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。
本发明还提供一种血清淀粉样蛋白A2作为分子标志物在制备检测肺癌试剂中的应用,所述血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。
本发明还提供一种试剂盒，以所述的血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。
优选的是，通过检测待测样品中血清淀粉样蛋白A2的表达量实现肺癌早期筛查和临床分期。
优选的是，所述待测样品为外周血样本。
本发明还提供一种检测所述的血清淀粉样蛋白A2的ELISA试剂盒，包括：血清淀粉样蛋白A2标准品、兔抗人WP02多克隆抗体包被的酶标板、辣根过氧化物酶标记的鼠抗人WM12单克隆抗体以及辅助试剂。
优选的是，所述辅助试剂包括辣根过氧化物酶标记的鼠抗人WM12单克隆抗体稀释液、待测样品稀释液、洗板液、显色液以及终止液。
进一步地，ELISA试剂盒包括：
ELISA板：已包被好纯化的兔抗人HE4多克隆抗体并已进行了封闭；
待检样品稀释液：含1％BSA和0.05％吐温-20的PBS；
酶标二抗：辣根过氧化物酶标记的鼠抗人HE4单克隆抗体，保存于含50％甘油的PBS；
酶标二抗稀释液：PH 7.2的PBS；
10×洗板液：0.2mol/L的Tris-HCl,1.5mol/L的NaCl,0.5％的吐温-20；
显色液：A液为0.05mol/L的柠檬酸和0.1mol/L的磷酸氢二钠，B液为含2mg/mL TMB的无水乙醇，C液为0.75％H2O2；
终止液：2M的硫酸；
阳性标准品：纯化的SAA2蛋白。
本发明公开了以下技术效果：
本发明公开了一种血清淀粉样蛋白A2作为分子标志物在制备检测肺癌试剂盒中的应用，通过ELISA试剂盒检测SAA2的表达量，发现SAA2基因表达与非小细胞肺癌(NSCLC)患者组织和血浆样本的密切相关性，首次揭示了SAA2基因高表达与肺癌临床分期密切关联，并开发了可用于临床的肺癌早期筛查和临床分期的快速检测试剂或试剂盒，实验验证，通过该试剂盒检测SAA2蛋白表达量检测非小细胞肺癌早期筛查和临床分期准确率达到68.89％，远高于现有技术中肺癌诊断的准确率，从而本发明为肺癌诊断提供新的手段。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为SAA2在人血浆中表达含量及其在非小细胞肺癌临床诊断和临床分期的应用；w：正常组和肺癌组中SAA2表达量分析结果；x：LUSC组和LUAD组中SAA2表达量分析结果；y：不同性别中SAA2表达量分析结果；z：正常组、LUSC组和LUAD组中不同性别对SAA2表达量的影响；α：肺癌不同分期中SAA2表达量的表达情况；β：ROC曲线分析ELISA试剂盒检测肺癌的临界值、灵敏度以及特异性。
具体实施方式
现详细说明本发明的多种示例性实施方式，该详细说明不应认为是对本发明的限制，而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式，并非用于限制本发明。另外，对于本发明中的数值范围，应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明，否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料，但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入，用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时，以本说明书的内容为准。
在不背离本发明的范围或精神的情况下，可对本发明说明书的具体实施方式做多种改进和变化，这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见的。本申请说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等，均为开放性的用语，即意指包含但不限于。
为实现本发明肺癌诊断技术，用于肺癌(具体为非小细胞肺癌)早期筛查和分期的检测，包括制备SAA2蛋白和SAA2抗体等，并用SAA2蛋白和SAA2抗体构建肺癌早期筛查和临床分期的试剂盒，具体包括以下步骤：
(1)重组蛋白制备：SAA2的编码序列(SEQ ID NO.1)通过RT-PCR获得，引物分别是：SAA2-F：5’-AACCGGCGCGCCAAGCTTCTCACGGGCCTGGTTTTCTGT-3’和SAA2-R：5’-TAGGTTAATTAACTACAGTTCAGCTGAAAATAAACTGATC-3’，模板用的是肺癌组织cDNA。PCR反应程序为：94℃预变性5min；94℃变性30s，55℃退火30s，72℃延伸30s，共30个循环；72℃延伸10min。
SEQ ID NO.1：
actataaatagcagccacctctccctggcagacagggacccgcagctcagctacagcacagatcagttatcctggggcatacagccataccattctgaaggtgtcttatctcctctgatctagagagcaccatgaagcttctcacgggcctggttttctgctccttggtcctgagtgtcagcagccgaagcttcttttcgttccttggcgaggcttttgatggggctcgggacatgtggagagcctactctgacatgagagaagccaattacatcggctcagacaaatacttccatgctcgggggaactatgatgctgccaaaaggggacctgggggtgcctgggctgcagaagtgatcagcaatgccagagagaatatccagagactcacaggccgtggtgcggaggactcgctggccgatcaggctgccaataaatggggcaggagtggcagagaccccaatcacttccgacctgctggcctgcctgagaaatactgagcttcctcttcactctgctctcaggagacctggctatgaggccctcggggcagggatacaaagttagtgaggtctatgtccagagaagctgagatatggcatataataggcatctaataaatgcttaagaggtggaa
经扩增获得的DNA片段经限制性内切酶BamHI/XohI双酶切后插入经BamHI/XohI双酶切的pGEX-4T-1中，构建重组质粒pGEX-4T-1-SAA2。转化重组质粒到BL21(DE3)中诱导表达，诱导表达的SAA2重组蛋白经过纯化，并通过SDS-PAGE检测。
(2)多克隆抗体制备：用生理盐水稀释SAA2重组蛋白，然后与弗氏不完全佐剂进行等比混合，吹打至抗原和佐剂完全混合形成稳定的乳剂，将该乳剂在兔子双肩周围的皮肤下进行皮下注射和后大腿肌肉注射。每次免疫前采血2mL(获得0.5-1mL免疫前血清)，并进行ELISA和Western Blotting检测。如果检测为阴性或抗体效价较低，则需要再次免疫。共免疫4次，末次放血20-30mL，纯化混合血样，平均每次纯化可以获得100-150mg抗体，最后进行ELISA和WB等质量检测。
(3)单克隆抗体制备：杂交瘤细胞株是以纯化的SAA2重组蛋白作为抗原筛选制备得到。获得能稳定分泌抗人SAA2蛋白的杂交瘤细胞株，并应用小鼠单克隆抗体亚型鉴定试剂盒鉴定单抗亚型。
(4)腹水制备及单抗纯化
用弗氏不完全佐剂0.5ml注射BALB/c小鼠腹腔。注射佐剂后一周将生长旺盛的杂交瘤细胞离心弃去培养基，用PBS或者不完全培养基重悬后注射小鼠腹腔，7-10天左右看到小鼠腹腔明显肿大，采集腹水12000rmp,离心5min收集上清。将腹水用0.06M醋酸钠(pH4.0)按1：3稀释，在4℃条件下搅拌加入正辛酸，一般按每毫升未稀释的腹水加入40uL正辛酸，然后4℃静置2h；4℃12000rmp，离心20min，可见正辛酸因温度低而从系统中结晶析出漂浮在腹水上层，弃去此结晶以及底部沉淀，向上清中加入1/10体积的10×PBS(0.1M，pH 7.4)，4℃条件下向其中加入硫酸铵固体至终浓度为0.277g/mL，静置1h以上；12000rmp，离心20min；弃上清，将沉淀溶于适量PBS中，再将其装入透析袋对PBS透析(透析液应为抗体体积的100倍以上，最好搅拌)，4℃过夜；12000rmp，离心3min，收集上清，紫外分光光度计测定纯化后的单克隆抗体含量，0.22um滤膜过滤后-80℃保存。
(5)辣根过氧化物酶标记纯化的单克隆抗体
用过碘酸盐氧化法获得酶标二抗：溶解5mg辣根过氧化物酶于1.2mL超纯水中，加入新配制的0.1mol/L的过碘酸钠0.3mL,室温静置20min,将上述溶液在1mmol/L的醋酸钠中4℃透析24h,期间每4h更换透析液。用0.02mol/L的碳酸钠溶液制备单抗样品至10mg/mL,将0.5mL制备好的抗体溶液加入经透析的辣根过氧化物酶溶液中，室温静置2h,在混合液中加入100μL硼氢化钠(4mg/mL的水溶液)，4℃反应2h，向混合液中加入等体积的饱和硫酸铵，4℃反应30min后12000rpm离心15min，弃上清，沉淀溶于1mL的PBS中，在PBS中透析过夜，次日12000rpm离心15min后取上清加入等体积的甘油，分装后-70℃保存。取少量酶标二抗，用纯化的SAA2作为包被抗原，间接ELISA方法检测。
(6)人肺癌标志物SAA2双抗体夹心ELISA检测方法的建立
应用获得的兔抗人SAA2多克隆抗体为捕获抗体,以辣根过氧化物酶标记的鼠抗人SAA2单克隆抗体作为检测抗体，建立检测肺癌标志物SAA2的多抗和单抗双抗体夹心ELISA方法。
肺癌分子标志物SAA2双抗体夹心ELISA试剂盒的包括：
(1)ELISA板：以兔抗人WP02多克隆抗体做为捕获抗体，以BSA/PBS(0.1M，pH 7.40)包被液稀释，100ul/孔，4℃下过夜包被(12-16h)；封闭备用。
(2)待检样品稀释液：含1％BSA和0.05％吐温-20的PBS溶液
(3)酶标二抗：辣根过氧化物酶标记的鼠抗人WM12单克隆抗体，保存于含50％甘油的PBS溶液；
(4)酶标二抗稀释液：pH 7.2的PBS溶液；
(5)10×洗板液：0.2mol/L的Tris-HCl,1.5mol/L的NaCl,0.5％的吐温-20；
(6)显色液：A液为0.05mol/L的柠檬酸和0.1mol/L的磷酸氢二钠，B液为含2mg/mLTMB的无水乙醇，C液为0.75％H2O2。
(7)终止液：2M的硫酸。
(8)阳性标准品：纯化的SAA2蛋白。
实施例1血清淀粉样蛋白A2在制备检测肺癌早期筛查和临床分期试剂盒中的应用
1、试验材料
收集90名肺癌患者和61名健康人的血样。所有将要用于ELISA检测的人外周血样本都在2小时内进行1600g，4℃离心15min，然后转移上清液到新的EP管中，做好标记，迅速放到-80℃冻存。
2、试验方法
从冰箱中取出血浆样品，并用磷酸盐缓冲盐水(PBS)稀释10倍。从冰箱中取出上述构建的ELISA试剂盒，放到实验台上使温度恢复室温，然后按照说明书稀释蛋白标准试剂和其他试剂。将样品按照每孔100μL加至96孔板中，在37℃下孵育1小时。接下来，每孔加入100μL检测溶液A，在37℃下孵育1小时。每孔加入检测溶液B 100μL，并于37℃孵育30min。每孔加入90μL稀释的底物溶液，并在37℃下孵育15min。最后，每孔加入50μL终止液，并立即在酶标仪(Thermo，USA)上于450nm条件下测量其OD值。
3、数据处理
用GraphPad Prism 8.0(GraphPad软件，LaJolla,CA,USA)对以上数据进行分析，用Student’s t检验(Student’s t test)分析是否有显著性差异。筛选差异基因和蛋白均在相应网站上操作。双侧t检验(double-tailed T test)用来分析两组之间的差异。单因素方差分析(ANOVA)用于3组及以上的数据分析，P<0.05表明有显著性差异。
4、数据总结
总共收集了151例血浆样本，他们的年龄分布在22到79岁之间，其中有90例被诊断为肺癌。
5、数据分析
SAA2的表达水平及诊断效能用ELISA检测后表明，与正常组相比，肺癌组中的SAA2显著上调(P<0.0001)(0.75±0.31VS 0.28±0.14，μg/mL；图1-w)。与正常组相比，LUAD组(p<0.0001)和LUSC组(p<0.001)SAA2明显上调；其中，LUSC比LUAD高(0.83±0.30VS 0.71±0.31，μg/mL；图1-x)，与基因数据库中SAA2的基因表达一致(图1)。此外，男性的SAA2高于女性(0.79±0.31VS 0.65±0.28；μg/mL；图1y)，LUSC的结果也是类似(0.76±0.32VS 0.65±0.28，μg/mL；图1z)。同时，值得注意的是，SAA2的表达随疾病分期的增加而增加(0.679±0.35VS0.683±0.15VS 0.78±0.30VS 0.81±0.31，μg/mL；图1α)。然后，ROC曲线分析显示其临界值为74.72，灵敏度为83.61％，特异性为91.11％。SAA2的AUC为0.9252，95％置信区间为0.8820至0.9685(P<0.0001，图1β)。最终，其准确性为68.89％。
由于肺腺癌(LUAD)和肺鳞癌(LUSC)是最常见的非小细胞肺癌类型，而上述的结果显示，SAA2蛋白可以作为检测非小细胞肺癌的分子标志物使用，并且利用ELISA法检测SAA2蛋白的表达量能够实现非小细胞肺癌早期筛查和临床分期，上述结果也是首次披露。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。
序列表
<110> 上海中乔新舟生物科技有限公司
<120> 血清淀粉样蛋白A2在制备检测肺癌试剂盒中的应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 637
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
actataaata gcagccacct ctccctggca gacagggacc cgcagctcag ctacagcaca 60
gatcagttat cctggggcat acagccatac cattctgaag gtgtcttatc tcctctgatc 120
tagagagcac catgaagctt ctcacgggcc tggttttctg ctccttggtc ctgagtgtca 180
gcagccgaag cttcttttcg ttccttggcg aggcttttga tggggctcgg gacatgtgga 240
gagcctactc tgacatgaga gaagccaatt acatcggctc agacaaatac ttccatgctc 300
gggggaacta tgatgctgcc aaaaggggac ctgggggtgc ctgggctgca gaagtgatca 360
gcaatgccag agagaatatc cagagactca caggccgtgg tgcggaggac tcgctggccg 420
atcaggctgc caataaatgg ggcaggagtg gcagagaccc caatcacttc cgacctgctg 480
gcctgcctga gaaatactga gcttcctctt cactctgctc tcaggagacc tggctatgag 540
gccctcgggg cagggataca aagttagtga ggtctatgtc cagagaagct gagatatggc 600
atataatagg catctaataa atgcttaaga ggtggaa 637
Claims (7)
1.一种血清淀粉样蛋白A2作为分子标志物在制备检测肺癌试剂盒中的应用,其特征在于，所述血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。
2.一种血清淀粉样蛋白A2作为分子标志物在制备检测肺癌试剂中的应用,其特征在于，所述血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。
3.一种试剂盒，其特征在于，以权利要求1或2中所述的血清淀粉样蛋白A2作为分子标志物对肺癌进行早期筛查和临床分期。
4.根据权利要求3所述的试剂盒，其特征在于，通过检测待测样品中血清淀粉样蛋白A2的表达量实现肺癌早期筛查和临床分期。
5.根据权利要求4所述的试剂盒，其特征在于，所述待测样品为外周血样本。
6.一种检测权利要求1或2中所述的血清淀粉样蛋白A2的ELISA试剂盒，其特征在于，包括：血清淀粉样蛋白A2标准品、兔抗人WP02多克隆抗体包被的酶标板、辣根过氧化物酶标记的鼠抗人WM12单克隆抗体以及辅助试剂。
7.根据权利要求6所述的检测血清淀粉样蛋白A2的ELISA试剂盒，其特征在于，所述辅助试剂包括辣根过氧化物酶标记的鼠抗人WM12单克隆抗体稀释液、待测样品稀释液、洗板液、显色液以及终止液。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110671159.6A CN113189335A (zh) | 2021-06-17 | 2021-06-17 | 血清淀粉样蛋白a2在制备检测肺癌试剂盒中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110671159.6A CN113189335A (zh) | 2021-06-17 | 2021-06-17 | 血清淀粉样蛋白a2在制备检测肺癌试剂盒中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113189335A true CN113189335A (zh) | 2021-07-30 |
Family
ID=76976477
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110671159.6A Pending CN113189335A (zh) | 2021-06-17 | 2021-06-17 | 血清淀粉样蛋白a2在制备检测肺癌试剂盒中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113189335A (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115932272A (zh) * | 2022-09-23 | 2023-04-07 | 上海市第十人民医院 | 血清学生物标志物在克罗恩病临床诊断中的应用 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6194163B1 (en) * | 1995-07-21 | 2001-02-27 | The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin | Method for the quantitative measurement of human acute phase serum amyloid A protein; recombinant protein; specific antibody |
| US20040072181A1 (en) * | 2002-01-22 | 2004-04-15 | Whitehead Alexander Steven | Methods for determining drug responsiveness |
| CN102243240A (zh) * | 2010-05-11 | 2011-11-16 | 中国科学院上海生命科学研究院 | 非小细胞肺癌分子标志物及其应用 |
| WO2012091465A2 (ko) * | 2010-12-31 | 2012-07-05 | (주)프로탄바이오 | 혈청 아밀로이드 ａ에 대한 단일클론항체 및 이를 생산하는 하이브리도마 세포 |
| CN111670363A (zh) * | 2017-12-19 | 2020-09-15 | 拜克门寇尔特公司 | 具有可变工作流的集成样品处理系统 |
-
2021
- 2021-06-17 CN CN202110671159.6A patent/CN113189335A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6194163B1 (en) * | 1995-07-21 | 2001-02-27 | The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin | Method for the quantitative measurement of human acute phase serum amyloid A protein; recombinant protein; specific antibody |
| US20040072181A1 (en) * | 2002-01-22 | 2004-04-15 | Whitehead Alexander Steven | Methods for determining drug responsiveness |
| CN102243240A (zh) * | 2010-05-11 | 2011-11-16 | 中国科学院上海生命科学研究院 | 非小细胞肺癌分子标志物及其应用 |
| WO2012091465A2 (ko) * | 2010-12-31 | 2012-07-05 | (주)프로탄바이오 | 혈청 아밀로이드 ａ에 대한 단일클론항체 및 이를 생산하는 하이브리도마 세포 |
| CN111670363A (zh) * | 2017-12-19 | 2020-09-15 | 拜克门寇尔特公司 | 具有可变工作流的集成样品处理系统 |
Non-Patent Citations (5)
| Title |
|---|
| CHRISTINE C. MCCORMACK 等: "Generation of soluble recombinant human acute phase serum amyloid A2 (A-SAA2) protein and its use in development of a A-SAA specific ELISA" * |
| YEOUN JIN KIM 等: "Quantification of SAA1 and SAA2 in lung cancer plasma using the isotype-specific PRM assays" * |
| 蒋敏 等: "肺癌组织与癌旁组织的差异蛋白谱" * |
| 覃慧婵: "应用iTRAQ标记联合LC-MS/MS技术筛选肺腺癌血浆差异蛋白的研究" * |
| 金旗: "肺血栓栓塞症蛋白质组学及介入治疗研究" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115932272A (zh) * | 2022-09-23 | 2023-04-07 | 上海市第十人民医院 | 血清学生物标志物在克罗恩病临床诊断中的应用 |
| CN115932272B (zh) * | 2022-09-23 | 2023-12-12 | 上海市第十人民医院 | 血清学生物标志物在克罗恩病临床诊断中的应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101160526B (zh) | 针对胃泌素释放肽前体的抗体及其应用 | |
| JP5670422B2 (ja) | 胃癌検出用マーカー及び胃癌検出方法 | |
| CN110187111B (zh) | 一种用于早期贲门癌筛查elisa试剂盒 | |
| CN113189335A (zh) | 血清淀粉样蛋白a2在制备检测肺癌试剂盒中的应用 | |
| CN107255711A (zh) | 骨桥蛋白用于制备或筛选慢加急性肝衰竭诊断试剂的用途 | |
| WO2015149450A1 (zh) | Ehd2抗体及其在制备乳腺癌免疫组化检测试剂中的应用 | |
| EP2863224A1 (en) | Kit for diagnosing malignant melanoma | |
| KR100832870B1 (ko) | 사스 코로나바이러스 뉴클레오캡시드 단백질에 대한 단클론항체 및 이것의 용도 | |
| CN111077312B (zh) | 一组肿瘤相关抗原在制备贲门癌早期筛查试剂盒中的应用 | |
| KR101777254B1 (ko) | En2 단백질을 특이적으로 인식하는 특정 항원으로부터 얻어진 단클론 항체 또는 이를 함유하는 전립선암 진단용 조성물 | |
| US20140363832A1 (en) | Molecular marker for the early detection of malignant pleural mesothelioma and the methods of its expression analysis using blood and pleural effusion samples | |
| JP5773979B2 (ja) | 胃癌検出用マーカー及び胃癌検出方法 | |
| US11746146B2 (en) | Antibody composition specifically recognizing an immunogenic fragment peptide of EN2 protein | |
| KR20120116518A (ko) | 폐암 조기 진단용 ＸＡＧＥ-1ａ 마커 및 이의 용도 | |
| CN106928352B (zh) | 一种抗psg3蛋白的单克隆抗体及其杂交瘤细胞株与应用 | |
| JPS61258173A (ja) | モノクロ−ナル抗体およびその製法ならびに用途 | |
| US20100221742A1 (en) | Novel cancer associated antibodies and their use in cancer diagnosis | |
| CN105277720B (zh) | 一种检测肝吸虫IgG4抗体的ELISA试剂盒及其制备方法 | |
| CN110865184B (zh) | Srsp蛋白和srsp抗原表位肽的应用及诊断和治疗肿瘤的产品 | |
| CN112574955B (zh) | 杂交瘤细胞lcz9h4及其分泌的单克隆抗体和应用 | |
| CN114874319B (zh) | 一种用于急性肾损伤检测的cryab抗体及其应用 | |
| WO2023245801A1 (zh) | 一种抗念珠菌甘露聚糖单克隆抗体及其应用 | |
| CN117004575A (zh) | 一种用于兔出血症病毒的杂交瘤细胞株及单克隆抗体和应用 | |
| CN114751961A (zh) | circ0005199-173aa蛋白及其在制备食管癌诊断产品中的应用 | |
| WO2017113565A1 (zh) | 基于蛋白标志物psg3辅助诊断肝癌或消化道癌症患者的试剂盒 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220323 Address after: 200436 room 1499, zone a, floor 5, building 1, No. 668, SHANGDA Road, Baoshan District, Shanghai Applicant after: Shanghai iselosen biomedical Co.,Ltd. Address before: 200439 room A0377, building 3, No. 112-118, Gaoyi Road, Baoshan District, Shanghai Applicant before: Shanghai Zhongqiao Xinzhou Biotechnology Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210730 |