CN107674115A - Folacin receptor alpha specific binding peptide 3 and its application - Google Patents
Folacin receptor alpha specific binding peptide 3 and its application Download PDFInfo
- Publication number
- CN107674115A CN107674115A CN201710649313.3A CN201710649313A CN107674115A CN 107674115 A CN107674115 A CN 107674115A CN 201710649313 A CN201710649313 A CN 201710649313A CN 107674115 A CN107674115 A CN 107674115A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- peptide
- folacin receptor
- application
- nucleotide sequence
- 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
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 41
- OVBPIULPVIDEAO-LBPRGKRZSA-N Folic acid Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 title claims abstract description 21
- 235000019152 folic acid Nutrition 0.000 title claims abstract description 20
- 239000011724 folic acid Substances 0.000 title claims abstract description 20
- 229940064302 folacin Drugs 0.000 title claims abstract description 19
- 230000009870 specific binding Effects 0.000 title description 8
- 229920001184 polypeptide Polymers 0.000 claims abstract description 16
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 16
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 10
- 210000004027 cell Anatomy 0.000 claims description 28
- 201000011510 cancer Diseases 0.000 claims description 4
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 238000001727 in vivo Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims 8
- 125000003729 nucleotide group Chemical group 0.000 claims 8
- 239000003814 drug Substances 0.000 claims 4
- 239000008194 pharmaceutical composition Substances 0.000 claims 4
- 239000003937 drug carrier Substances 0.000 claims 2
- 102000009027 Albumins Human genes 0.000 claims 1
- 108010088751 Albumins Proteins 0.000 claims 1
- 208000030808 Clear cell renal carcinoma Diseases 0.000 claims 1
- 206010009944 Colon cancer Diseases 0.000 claims 1
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 claims 1
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 claims 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims 1
- 230000021736 acetylation Effects 0.000 claims 1
- 238000006640 acetylation reaction Methods 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 150000001408 amides Chemical group 0.000 claims 1
- 210000004899 c-terminal region Anatomy 0.000 claims 1
- 239000003153 chemical reaction reagent Substances 0.000 claims 1
- 206010073251 clear cell renal cell carcinoma Diseases 0.000 claims 1
- 208000029742 colonic neoplasm Diseases 0.000 claims 1
- 239000000470 constituent Substances 0.000 claims 1
- 229940079593 drug Drugs 0.000 claims 1
- 201000003908 endometrial adenocarcinoma Diseases 0.000 claims 1
- 208000029382 endometrium adenocarcinoma Diseases 0.000 claims 1
- 150000004665 fatty acids Chemical group 0.000 claims 1
- 239000003446 ligand Substances 0.000 claims 1
- 210000005075 mammary gland Anatomy 0.000 claims 1
- 230000004048 modification Effects 0.000 claims 1
- 238000012986 modification Methods 0.000 claims 1
- 239000003068 molecular probe Substances 0.000 claims 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 claims 1
- 102000039446 nucleic acids Human genes 0.000 claims 1
- 108020004707 nucleic acids Proteins 0.000 claims 1
- 150000007523 nucleic acids Chemical class 0.000 claims 1
- 239000000523 sample Substances 0.000 claims 1
- 210000002966 serum Anatomy 0.000 claims 1
- 210000001768 subcellular fraction Anatomy 0.000 claims 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 claims 1
- 230000014509 gene expression Effects 0.000 abstract description 12
- 238000002823 phage display Methods 0.000 abstract description 6
- 210000004881 tumor cell Anatomy 0.000 abstract description 4
- 238000002626 targeted therapy Methods 0.000 abstract description 2
- 238000009509 drug development Methods 0.000 abstract 1
- 238000003384 imaging method Methods 0.000 abstract 1
- 239000002547 new drug Substances 0.000 abstract 1
- 241001515965 unidentified phage Species 0.000 description 35
- 238000002965 ELISA Methods 0.000 description 25
- 239000007788 liquid Substances 0.000 description 23
- 239000006228 supernatant Substances 0.000 description 22
- 239000000243 solution Substances 0.000 description 17
- 238000005406 washing Methods 0.000 description 15
- 238000012216 screening Methods 0.000 description 14
- 102000005962 receptors Human genes 0.000 description 13
- 108020003175 receptors Proteins 0.000 description 13
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 12
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 12
- 239000007853 buffer solution Substances 0.000 description 12
- 238000010790 dilution Methods 0.000 description 12
- 239000012895 dilution Substances 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 239000006180 TBST buffer Substances 0.000 description 9
- 238000005119 centrifugation Methods 0.000 description 8
- 238000012163 sequencing technique Methods 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 230000027455 binding Effects 0.000 description 6
- 238000009739 binding Methods 0.000 description 6
- 238000000576 coating method Methods 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000011248 coating agent Substances 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- PXFBZOLANLWPMH-UHFFFAOYSA-N 16-Epiaffinine Natural products C1C(C2=CC=CC=C2N2)=C2C(=O)CC2C(=CC)CN(C)C1C2CO PXFBZOLANLWPMH-UHFFFAOYSA-N 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 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 3
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 3
- 108010067902 Peptide Library Proteins 0.000 description 3
- 206010057249 Phagocytosis Diseases 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000008782 phagocytosis Effects 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000004042 decolorization Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- 101710192393 Attachment protein G3P Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 101100298998 Caenorhabditis elegans pbs-3 gene Proteins 0.000 description 1
- 108090000565 Capsid Proteins Proteins 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 102000000844 Cell Surface Receptors Human genes 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 102100023321 Ceruloplasmin Human genes 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 241001524679 Escherichia virus M13 Species 0.000 description 1
- 102000010451 Folate receptor alpha Human genes 0.000 description 1
- 108050001931 Folate receptor alpha Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000005267 amalgamation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 102000006815 folate receptor Human genes 0.000 description 1
- 108020005243 folate receptor Proteins 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 125000003929 folic acid group Chemical group 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000007973 glycine-HCl buffer Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000009149 molecular binding Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920006324 polyoxymethylene Polymers 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57415—Specifically defined cancers of breast
-
- 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
- G01N33/57419—Specifically defined cancers of colon
-
- 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
- G01N33/57442—Specifically defined cancers of the uterus and endometrial
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2410/00—Assays, e.g. immunoassays or enzyme assays, involving peptides of less than 20 animo acids
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- General Physics & Mathematics (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Reproductive Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Inorganic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The small peptide combined the present invention relates to a kind of folacin receptor alpha specific and its application, belong to biological technical field.The peptide sequence of the present invention is SGVYKVAYDWQH.This polypeptide is obtained by being screened from phage display library, can be with expression folacin receptor α tumour cell specific bond, and targeted therapy and tumor imaging for tumour provide a kind of new targeted molecular, the potential value with new drug development.
Description
Technical field:
The invention belongs to biological technical field, is related to a kind of small peptide of folacin receptor alpha specific combination and its in tumor target
Application in.
Background technology:
Display technique of bacteriophage is that allogenic polypeptide or certain of albumen and bacteriophage capsid protein are carried out into amalgamation and expression, is made
Foreign protein can be illustrated in surfaces of viral particles, while the DNA of encoding foreign proteins is located in the virion.Random 12
Peptide phage display library is that random dodecapeptides are fused on M13 phage minor coat proteins (pIII), for a combination
Library.N-terminal of the dodecapeptide expression shown in pIII.This is just between substantial amounts of rondom polypeptide and its DNA encoding sequence
Direct bridge is established, then target is obtained to carry out in-vitro screening with various target molecules (antibody, enzyme, cell surface receptor etc.)
Molecule binding peptide.External option program is exactly that phage library is incubated altogether with solid phase target molecule in simple terms, and washing removes uncombined
Bacteriophage, then afford the bacteriophage that can be specifically bound with target molecule.The bacteriophage being eluted also needs to be expanded, and enters
Combination/amplification cycles of row next round, to be enriched with specific binding bacteriophage., can by DNA sequencing after 3~4 wheels " elutriation "
To obtain the peptide sequence of each specific binding.
Folacin receptor (folate receptor, FR) is to combine and transport folic acid and its derivative into the important of cell
Transporter, mainly exist in vivo with three kinds of hypotypes:FR α, β and γ.Folacin receptor α (folate receptor α, FR α) is one
Kind is anchored to the glycoprotein of cell membrane surface by glycolsyl-phosphatidylinositol (GPI).Existing document report, oophoroma, lung cancer, liver
FR α express in high in the tissue such as cancer, breast cancer, and restricted expression in the normal tissue, therefore are considered as great potential
Ovary carcinoma marker or tumor associated antigen (TAA);The high degree of specificity that FR α have to oophoroma, treatment can be used as related swollen
The target spot of knurl;Part research also indicates that early diagnosis of the FR α for oophoroma has important value.Based on this, if can obtain with
FR α have the short peptide sequence of higher affinity, will provide new approaches for the magnetic target therapy of cancer and diagnosis.
The content of the invention
Goal of the invention
It is an object of the invention to using display technique of bacteriophage obtain it is a kind of can be combined with folacin receptor alpha specific it is more
Peptide.The polypeptide can be used for the positive tumour cell of targeting folacin receptor alpha expression.
Technical scheme
To achieve these goals, the technical solution adopted in the present invention is as follows:
1) the affine screening of phage display peptide library:Using folacin receptor α recombinant proteins as target, with phage display ten
Dipeptides storehouse carries out four-wheel biological screening.The selective mechanisms rate of recovery and polyclonal ELISA are often taken turns, to judge whether screening is effective;It is logical
Cross each clone of monoclonal ELISA detections and the binding ability of folacin receptor α recombinant proteins;
2) bacteriophage positive monoclonal DNA preparation and sequencing identification:According to above-mentioned ELISA results, select positive value high
Bacteriophage monoclonal, after being expanded to it carry out sequencer module fast purifying surveyed with producing sufficiently pure template
Sequence, it is sent to company from -96gIII sequencing primers and is sequenced, analysis positive bacteriophage accordingly shows peptide sequence;
3) cell ELISA detection phage clone and native cell surface folacin receptor α combination:Choose FR alpha expressions sun
Property Cell line SKOV3, and using FR alpha expressions negative cells strain HepG2 as control, pass through ELISA and detect phage clone and cell
The combination of strain;
4) phage clone and FR alpha expression positive cell strains SKOV3 combination are further confirmed that by flow cytometry.
Beneficial effect
FR alpha bindings provided by the invention, FR α recombinant proteins can be specifically bound, while pass through cell ELISA and streaming
Cytometry experiment is analyzed, and the phage display small peptide for finding to filter out can be combined with the natural FR alpha specifics of cell surface, is had latent
Medical science and pharmacy value.
Brief description of the drawings:
The polyclonal ELISA of Fig. 1;
Fig. 2 positive monoclonals and the research of folacin receptor α recombinant protein binding abilities.Fig. 2A monoclonal phages
ELISA screening positive clones (clone irised out is the displaying of folacin receptor alpha specific binding peptide 3 clone);Fig. 2 B. positive monoclonals
The further checking combined with folacin receptor α recombinant proteins;
The combination of positive monoclonal and FR alpha expression positive cells SKOV3 that the detection of Fig. 3 cell ELISAs screens, with M13
And FR alpha expression negative cells HepG2 is control;
Fig. 4 flow cytometry tests analyze positive monoclonal and SKOV3 specific binding situation, in control group such as figure
Shown in dotted line, the SKOV3 cells of anti-M13 antibody and FITC fluorescence antibodies are only added.
Embodiment:
In order to more clearly illustrate the present invention, the description below provides more specific embodiment, the technology of this area
Personnel are it should be understood that the present invention is not merely defined in following embodiments.
The affine screening of the phage display peptide library of embodiment 1
Random dodecapeptides phage display library is purchased from NEB companies, 100 μ l, and titre is 1.5 × 1013pfu/ml.It is stored in
In TBS buffer solutions (50mM Tris-HCl, 150mM NaCl [PH7.5]) containing 50% glycerine, storage capacity 2.7 × 109Individual conversion
Son.Escherichia coil ER2738 are the Host Strains in this peptide storehouse.
(1) preparation
FR α recombinant proteins are diluted to final concentration 50 μ g/ml with coating buffer solution (carbonate buffer solution [PH9.6]), it is dilute
Solution after releasing is rotated until surface moistens completely repeatedly with every μ l coated elisa plates of hole 100.4 DEG C were incubated in wet box
Night.
(2) affine screening
Second day coating buffer outwelled in ELISA Plate, with TBST buffer solutions (TBS+ volume ratios 0.1% [v/v] Tween-20)
Board-washing, incline buffer solution, claps and is got rid of to remove remaining liquid on clean blotting paper.200 μ l envelopes are added in every hole afterwards
Blocking solution (0.1M NaHCO3, 5mg/ml BSA, 0.02%NaN3[PH8.6]), put 4 DEG C of overnight incubations or 37 DEG C of incubations in wet box
At least 1 hour.Discard and blockade liquid, TBST buffer solutions are filled it up with per hole, board-washing 6 times, washing 5 minutes, are placed on decolorization swinging table every time
Carry out.Incline buffer solution again, claps and is got rid of to remove remaining liquid on clean blotting paper, performs quick during this experimental procedure
To avoid orifice plate from drying.Primary libraries are diluted with TBST buffer solutions, take the μ l of library liquid 100 after dilution to be added to coating FR in advance
In the ELISA Plate micropore of α recombinant proteins, the bacteriophage quantity of addition is about 5 × 1012.In wet box 4 DEG C be incubated overnight or 37 DEG C
Place 2 hours, discard liquid in hole, removing residual solution is got rid of into ELISA Plate inversion bat, and with TBST buffer solutions board-washing 10 times, grasp
Make the same (washing away uncombined bacteriophage).After last time washing, liquid in hole is clapped and dried only, is added into ELISA Plate micropore
It is bound to elute to enter 100 μ l non-specificity buffer solution such as 0.2M glycine-HCIs Glycine-HCl buffer solutions [PH2.2]
Bacteriophage, it is gentle on decolorization swinging table to shake 10 minutes fully to elute under room temperature condition, then with 15 μ l 1M Tris-HCl
[PH9.1] neutralizes above-mentioned eluent, is transferred in a clean EP pipe.Routinely M13 methods take 5 μ l eluents to be used for determining
The titre of eluate, remaining eluent are added to 20ml and are in progress first round bacteriophage in the ER2738 strains of logarithm early stage
The amplification of eluate, in 37 DEG C, 220rpm shaking table cultures 4.5 hours, amplified production is transferred in clean centrifuge tube, through 4 DEG C,
After 10, the 000rpm operations of centrifugation 10 minutes, about 80% bacteriophage supernatant is taken, is transferred in another clean centrifuge tube, adds 1/6
The PEG/NaCl (20% [w/v] PEG-800,2.5M NaCl) of volume, 4 DEG C are placed at least 1 hour or handled overnight, are bitten
Bacterial sediment.4 DEG C, 10,000rpm centrifugation 10 minutes, discard remaining supernatant with micropipettor, can carry out again it is of short duration centrifugation with
Thoroughly inhale and abandon supernatant.Sediment is resuspended in 200 μ l TBS buffer solutions, is taken supernatant after centrifuging again, is transferred to clean centrifugation
Guan Zhong, this is the eluate after expanding.5 μ l eluates are taken to carry out the measure of phage titre, remainder is used for the second wheel
Affine screening.Repeat above step and carry out four-wheel screening altogether.The washing times of TBST in increase washing step are screened by wheel.
(3) measure of phage titre
ER2738 strains are inoculated with 10ml LB culture mediums, 37 DEG C of shaking table cultures to mid-log phase (OD600It is left 0.5
It is right).Period dissolves by heating top-layer agar in micro-wave oven, is dispensed into 5ml sterilizing EP pipes, often pipe 3ml, pipe are several according to phagocytosis
Depending on body dilution gradient, each pipe of dilution gradient one.It is standby that the EP pipes of packing are placed in 45 DEG C of water-baths;Prepare LB/ simultaneously
IPTG/Xgal culture plates, it is standby to be placed in 37 DEG C of incubators.10 times of gradients are carried out with LB culture mediums to the bacteriophage supernatant of collection
Dilute (the bacteriophage dilution range typically expanded:108~1011).By the ER2738 bacterium solutions in mid-log phase after the completion of dilution
It is dispensed into several 1.5ml sterilizing EP pipes, often the μ l of pipe 200.10 μ l are taken to add immediately the dilution of each gradient diluted
Into the EP pipes containing bacterium solution, vibration mixes to be incubated 5 minutes after 37 DEG C.Then the top-layer agar of 45 DEG C of placements is taken out, Guan Zhong
Content is transferred completely into top-layer agar immediately, and the LB/IPTG/Xgal cultures that pre-temperature is crossed are poured over after quick reverse mixing
On flat board, gently rocking flat board is uniformly distributed top-layer agar.After solidification a period of time to be cooled, 37 DEG C of inversion overnight incubations.
Flat board of the plaque sum at 100 or so is chosen, the plaque number to be grown on flat board is counted and calculates bacteriophage effect
Valency (pfu).According to the phagocytosis scale of construction (input titre Input) of every wheel input screening and the phagocytosis scale of construction afforded, (output is dripped
Spend Output), the output/input ratio often taken turns can be calculated, reflects the enrichment degree (rate of recovery Recovery) of specific bacteriophage.
Screen and find by four-wheel, having obtained effective enrichment with the bacteriophage of the high-affinity of FR α recombinant proteins specific binding (is shown in Table
1)。
Table 1 often takes turns the situation of input titre, output titre and the rate of recovery
The polyclonal ELISA identifications of the bacteriophage of embodiment 2
FR α recombinant proteins are diluted to the μ g/ml of final concentration 10 with carbonate buffer solution [PH9.6], the μ l of coating 100 per hole, 4
DEG C coating overnight.Molecule solution is discarded every other day, and is clapped on clean blotting paper and gets rid of removal residual liquid, and button is dry, is delayed with TBS
After fliud flushing washed once, 200 μ l are added per hole and blockade liquid, 37 DEG C are placed 1~2 hour.Throw away and blockade liquid, TBS buffer solutions washing 3
It is secondary, 5 minutes every time, firmly patted on clean blotting paper, get rid of net cleaning solution.Wash and 100 μ l dilutions are added in every hole
Each round screening amplification after eluent (i.e. bacteriophage supernatant), 37 DEG C of incubations discard liquid in hole after 2 hours, and use TBST
Buffer solution and TBS buffer solutions wash 3 times successively, 5 minutes every time, clap and add the anti-M13 of mouse that HRP is marked after getting rid of removing cleaning solution
As secondary antibody (being diluted to working concentration with liquid is blockaded), 37 DEG C are incubated 2 hours antibody, discard liquid in hole, washing operation is same as above
One operation.Add 100 μ l tmb substrate liquid per hole to be developed the color, room temperature lucifuge is incubated 10 minutes, and liquid should be by without discoloration in hole
For blueness;50 μ l 1M H are added per hole2SO4Terminate liquid, color development stopping reaction, detects OD on ELIASA450Value.
As a result show, as number of screening round increases, the affinity that eluate is combined with FR α recombinant proteins is stepped up, but
In fourth round, affinity has declined (see Fig. 1).
The ELISA identifications and sequencing identification of the bacteriophage monoclonal of embodiment 3
(1) ELISA detects binding ability of the monoclonal phage displaying small peptide to target molecule
The acquisition of bacteriophage to be measured:ER2738 strains need to be inoculated with advance, 37 DEG C of cultures to logarithm early stage, by this logarithm early stage
Culture is transferred in 2ml deep-well plates, per the μ l of hole 600.It is used to detecting the flat board of titre that (total amount is 100 in selection four-wheel screening
The flat board of individual or so and following plaque), with the multiple blue plaques of the random picking of pipette tips into deep-well plates, 37 DEG C,
220rpm shaking table cultures 4.5 hours.4 DEG C, 10,000rpm centrifugations 10 minutes, precipitation is discarded, takes 100 μ l supernatants to be used for per hole
ELISA is detected, and remaining supernatant collects temporary 4 DEG C of preservations.
ELISA is detected:Target molecule FR α recombinant proteins are dissolved in by carbonate buffer solution [PH9.6] with the μ g/ml of final concentration 10
In, 100 μ l are coated with per hole, 4 DEG C of coatings are stayed overnight in the wet box of sealing.Unnecessary molecule solution is thrown away, and in clean paper handkerchief
After the unnecessary liquid of removing is got rid of in upper bat, 200 μ l confining liquids are added per hole, 37 DEG C are closed 1~2 hour.Throw away and blockade liquid, TBST washings
6 times, clap after drying net ELISA Plate, the 100 μ l supernatants obtained in deep-well plates are sequentially added in order, room temperature reaction 1~2 is small
When, then 6 times (operation is same as above) are washed with TBST, with the anti-M13 antibody blockaded liquid and marked by 1: 10000 dilution proportion HRP, by every
In the μ l adding holes of hole 100, react at room temperature 1~2 hour, TBST fully washs 6 times (step is same as above), and 100 μ l TMB are added per hole
(now with the current) colour developing of substrate solution, lucifuge add 50 μ l 1M H in every hole after acting on 10 minutes2SO4Terminate liquid, color development stopping are anti-
Should, OD is detected on ELIASA450Value.
As a result Fig. 2 is seen.By Preliminary detection result, (clone that Fig. 2A is irised out is folacin receptor alpha specific binding peptide 3 to Fig. 2A
Displaying clone), independent picking clone, by its binding ability (figure with FR α recombinant proteins of further ELISA experimental verifications
2B), wherein PBS is as blank control, and there were significant differences compared with PBS control group for experimental group (P < 0.05).
(2) bacteriophage positive monoclonal DNA preparation and sequencing identification
According to above-mentioned ELISA results, choose after positive bacteriophage monoclonal is expanded and carry out the quick of sequencer module again
Purifying is sequenced with producing sufficiently pure template:ER2738 overnight cultures are inoculated in LB culture mediums according to 1: 100 and shaken
After shaking logarithmic phase, 1ml is into culture tube for packing.10 μ l bacteriophages positive colonies are taken into above-mentioned 1ml culture tubes.37 DEG C of shaking tables
Culture 4.5~5 hours.Culture is transferred in microcentrifugal tube, is centrifuged 30 seconds.Supernatant is transferred to a clean centrifuge tube, and this is
Bacteriophage reservoir is expanded, can temporary 4 DEG C of storages if not carrying out subsequent experiment immediately.
The fast purifying of sequencing template:(the volume about 1ml) containing bacteriophage supernatant of above-mentioned collection is taken, adds 400 μ l PEG/
NaCl, overturn and mix, 12,000rpm is centrifuged 10 minutes after room temperature is placed 10 minutes, is abandoned supernatant, can be carried out centrifugally operated again, thorough
Bottom, which is inhaled, abandons remaining supernatant.200 μ l iodide buffer solutions (10mM Tris-HCl, 1mM EDTA, 4M NaI is added in sediment
[PH8.0]) and sediment is thoroughly resuspended, 500 μ l ethanol are added, are incubated at room temperature 10 minutes (precipitating DNA);12,000rpm
Centrifugation 10 minutes, abandons supernatant.12,000rpm is centrifuged 10 minutes after cleaning precipitation once with 70% ethanol of 1ml precoolings, abandons supernatant
After uncap and air-dry overnight, ethanol is fully volatilized.Finally precipitation is resuspended in 30 μ l distilled waters, this is sequencing template, takes 5
μ l are detected with 1% agarose gel electrophoresis, and the DNA selection -96gIII sequencing primers after detection is qualified are sent to company and are sequenced,
Analyze corresponding 12 peptide sequence of bacteriophage.
By comparing, applicant obtains a small peptide SEQ ID NO.1:SGVYKVAYDWQH.
The cell ELISA of embodiment 4 detects the combination of phage display small peptide and cell
To verify the combination situation of FR α under the native state of the polypeptide screened and cell surface expression, high table is chosen
Up to FR α ovarian cancer cell SKOV3, and unrelated hepatocellular carcinoma H22 is set to carry out cell ELISA for control.SKOV3 is thin
Born of the same parents and HepG2 cell culture are to cell density up to more than 80%, and observation cellular morphology is good under the microscope, through 0.25% pancreatin
Add corresponding fresh culture after digestion to be resuspended and carry out viable count, adjustment cell density to 2 × 105Individual/ml, by every
The μ l of hole 100 spread 96 porocyte culture plates, 37 DEG C of overnight incubations.Every other day inhale abandon supernatant, with sterile PBS buffer (137mM NaCl,
2.7mM KCl, 10mM Na2HPO4, 2mM KH2PO4[PH7.4]) softly wash twice, 5 minutes every time, at room temperature with more than 10%
Polyformaldehyde fixes 15 minutes, and fixer is abandoned in suction, and with the soft board-washing of PBS 3 times, 5 minutes every time, with 5% skim milk
(PBS dilutions) is closed 2 hours for 37 DEG C as confining liquid.Confining liquid is abandoned, after PBS can do simple rinsing, corresponding bacteriophage is fitted
Added after dilution in Tissue Culture Plate, and one group of negative control (wild type M13) is set, per the μ l of hole 100,4 DEG C of overnight incubations.
Inhale every other day and abandon supernatant, and 3 times are washed successively with PBST (PBS+ volume ratios 0.1% [v/v] Tween-20), PBS, every time
5 minutes.The anti-M13 antibody for being diluted to the HRP of working concentration (1: 10000) with confining liquid and being marked is added, per the μ l of hole 100,37
DEG C it is incubated 2 hours, after ibid carrying out washing operation, chromogenic reaction and terminating reaction are the same as Phage-ELISA step.
As a result such as Fig. 3.Compared with wild type M13, experimental group polypeptide and SKOV3 have a preferable combination, and compared to
HepG2 combination, there is obvious specific (P < 0.05), illustrate FR alpha bindings and tumor cell surface table that screening obtains
The natural FR α reached can be specifically bound.
The combination situation of the Flow cytometry phage display small peptide of embodiment 5 and cell surface FR α
Experiment purpose is tested with cell ELISA.The positive bacteriophage monoclonal screened using Flow cytometry
Combination situation to SKOV3 tumour cells.
Cell processing:To cell density up to more than 80%, micro- Microscopic observation cellular morphology is good for SKOV3 cell culture,
Digested through 0.25% pancreatin, add fresh culture and terminate digestion, 1,500rpm centrifugation 5 minutes, PBS resuspensions prepare slender
Born of the same parents' suspension, carry out viable count, adjustment cell concentration to 1~2 × 106Individual/ml, it is dispensed into 1.5ml EP pipes, often pipe 250
The μ l of μ l~350.After 3,000rpm centrifugations 5 minutes, the bacteriophage supernatant that 100 μ l of often pipe addition have diluted is as primary antibody (dilution
For 2%FBS-PBS), the bacteriophage supernatant corresponding with the polypeptide that sequencing obtains is selected, after 4 DEG C are incubated 1 hour, takes out 3,
000rpm is centrifuged 5 minutes, abandons supernatant, and often pipe plus 1ml PBS are washed 2~3 times, and supernatant is abandoned in suction.Add the mouse source that 100 μ l have diluted
Anti- M13 secondary antibodies, 4 DEG C are placed 1 hour, are taken out ibid operation and are washed, are eventually adding the sheep anti mouse fluorescence two with FITC labels
It is anti-, by 1: 500 thinner ratio (2%FBS-PBS dilutions), per the μ l of hole 100, notice that this step needs lucifuge to operate.4 DEG C of lucifuges are incubated 1
Hour, washing step is same as above, and last PBS for adding appropriate volume according to cell precipitation after having washed is resuspended, and up flow type is thin
Born of the same parents' instrument is detected.
As a result show, see Fig. 4, the cell line that bacteriophage supernatant can be preferably with the high expression of FR α specifically binds, tied
Conjunction rate is 31.6%.
Claims (12)
1. a kind of polypeptide for targetting folacin receptor α, it is characterised in that amino acid sequence is SEQ ID NO.1:Ser-Gly-Val-
Tyr-Lys-Val-Ala-Tyr-Asp-Trp-Gln-His。
2. a kind of polypeptide for targetting folacin receptor α, it is characterised in that for the derivative of polypeptide described in claim 1, its derivative
One is connected to including but not limited to acetylation modification thing, fatty acid chain trim, PEG trims, the motif both ends of polypeptide
Individual Cys, polypeptide head and the tail acid amides ring.
3. a kind of fused polypeptide or albumen, it is characterised in that the N-terminal or C-terminal or tundish 1-2 containing claim of more peptide or proteins
Described peptide sequence, its merge part including but not limited to human serum albumins (HSA), Fc fragments, antibody or antibody fragment,
Go back to the nest molecule, targeted molecular, affinity ligand, cell penetrate peptide, in vivo escape molecule, subcellular fraction targeted molecular, core targeted molecular
Or their conjugate and mixture.
4. encode the nucleotide sequence of polypeptide described in claim 1.
5. a kind of nucleotide sequence, it is characterised in that the both ends of nucleic acid molecules or tundish are containing the nucleotides sequence described in claim 4
Row.
6. a kind of pharmaceutical composition, it is characterised in that lead to comprising the peptide sequence described in claim 1-3 with active constituents of medicine
Cross and be covalently or non-covalently coupled, or drug carrier is passed comprising the polypeptide described in claim 1-3.
7. a kind of molecular probe, it is characterised in that the probe includes the polypeptide described in claim 1-3.
8. a kind of pharmaceutical composition, it is characterised in that include the nucleotide sequence described in claim 4-5 and other nucleotides sequences
Row, which pass through, to be covalently or non-covalently coupled, or passs drug carrier comprising the nucleotide sequence described in claim 4-5.
9. claim 1-3 peptide sequence, claim 4-5 nucleotide sequence or claim 6-8 pharmaceutical composition
Application in medicine is prepared.
10. claim 1-3 peptide sequence, claim 4-5 nucleotide sequence or claim 6-8 pharmaceutical composition
In the application of tumour diagnostic reagent.
11. according to claim 8-10 application, it is characterised in that the medicine is preferred for tumor-targeting drug.
12. according to claim 8-10 application, it is characterised in that the tumour, which includes folacin receptor α, what is expressed to a certain degree
Benign or malignant tumour, such as oophoroma, adenocarcinoma of endometrium, non-small cell lung cancer, clear cell carcinoma of kidney, colon cancer and mammary gland
Cancer.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710649313.3A CN107674115A (en) | 2017-07-28 | 2017-07-28 | Folacin receptor alpha specific binding peptide 3 and its application |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710649313.3A CN107674115A (en) | 2017-07-28 | 2017-07-28 | Folacin receptor alpha specific binding peptide 3 and its application |
Publications (1)
Publication Number | Publication Date |
---|---|
CN107674115A true CN107674115A (en) | 2018-02-09 |
Family
ID=61134248
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710649313.3A Pending CN107674115A (en) | 2017-07-28 | 2017-07-28 | Folacin receptor alpha specific binding peptide 3 and its application |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107674115A (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105039333A (en) * | 2015-08-21 | 2015-11-11 | 天津医科大学 | Liver cancer targeted peptide and application thereof |
-
2017
- 2017-07-28 CN CN201710649313.3A patent/CN107674115A/en active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105039333A (en) * | 2015-08-21 | 2015-11-11 | 天津医科大学 | Liver cancer targeted peptide and application thereof |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112010966B (en) | Monoclonal antibody aiming at non-RBD (radial basis function) region of new coronavirus spike protein and application thereof | |
US7211395B2 (en) | Serum albumin binding moieties | |
CN110144009A (en) | CD47 single domain antibody and application thereof | |
CN109096395A (en) | Blocking-up type CD47 nano antibody and application thereof | |
JPH10506463A (en) | Antigen-binding peptide (abtide) from peptide library | |
CN111205351A (en) | PD-1 targeted blocking peptide and application thereof | |
CN107344968A (en) | A kind of time-resolved fluorescence immunoassay method for being used to detect avian influenza virus H7N9 | |
CN108892723B (en) | Single-domain heavy chain antibody for detecting porcine epidemic diarrhea virus, preparation method and application | |
CN108017712A (en) | A kind of peptide backbone of shark antibody sources and its application | |
KR20140080504A (en) | Screening methods and uses thereof | |
CN107353325A (en) | Folacin receptor alpha specific WHWTNWGKTSPA and its application | |
US20230029322A1 (en) | Tools and methods to detect and isolate colibactin producing bacteria | |
CN107446021A (en) | Folacin receptor alpha specific binding peptide 5 and its application | |
CN107793471A (en) | Folacin receptor alpha specific binding peptide 4 and its application | |
WO2022262509A1 (en) | Leptin receptor affinity peptide and use thereof | |
CN107446020B (en) | Folacin receptor alpha specific peptide 2 and its application | |
CN107674115A (en) | Folacin receptor alpha specific binding peptide 3 and its application | |
CN114773460B (en) | Nano antibody of targeted rotavirus protein and application thereof | |
CN113999308B (en) | Nano antibody targeting cadherin 17 and application thereof | |
CN106967154B (en) | Screening method of human colon cancer cell specific binding polypeptide and application thereof | |
CN109503711A (en) | A kind of dual-functional nanometer antibody, encoding gene and its application for blood clotting method detection PCV2 virus | |
CN105037499A (en) | Method utilizing phage antibody library to screen human histamine receptor 4 (HR4) epitope mimic peptide and vaccine construction method thereof | |
CN105017385A (en) | Vaccine based on simulation of histamine acceptor 4 (HR4) epitope and construction method thereof | |
CN108822189A (en) | A kind of targeting combines specific polypeptide and its application of lymphocytic cancer cell system | |
JP2008515419A (en) | Methods for screening antibody libraries |
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 | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180209 |