CA2392075C - Method for the preparation of purified hcv rna by exosome separation - Google Patents
Method for the preparation of purified hcv rna by exosome separation Download PDFInfo
- Publication number
- CA2392075C CA2392075C CA2392075A CA2392075A CA2392075C CA 2392075 C CA2392075 C CA 2392075C CA 2392075 A CA2392075 A CA 2392075A CA 2392075 A CA2392075 A CA 2392075A CA 2392075 C CA2392075 C CA 2392075C
- Authority
- CA
- Canada
- Prior art keywords
- hcv
- exosome
- preparation
- plasma
- rna
- 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.)
- Expired - Lifetime
Links
- 210000001808 exosome Anatomy 0.000 title claims abstract description 73
- 238000000034 method Methods 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims description 23
- 238000000926 separation method Methods 0.000 title abstract description 9
- 241000711549 Hepacivirus C Species 0.000 claims abstract description 60
- 239000002245 particle Substances 0.000 claims abstract description 34
- 210000002381 plasma Anatomy 0.000 claims abstract description 26
- 210000004027 cell Anatomy 0.000 claims description 23
- 239000011324 bead Substances 0.000 claims description 22
- 238000004113 cell culture Methods 0.000 claims description 13
- 239000006228 supernatant Substances 0.000 claims description 13
- 238000001085 differential centrifugation Methods 0.000 claims description 11
- 108090000623 proteins and genes Proteins 0.000 claims description 9
- 102000004169 proteins and genes Human genes 0.000 claims description 8
- 239000012228 culture supernatant Substances 0.000 claims description 7
- 230000001413 cellular effect Effects 0.000 claims description 5
- 239000000872 buffer Substances 0.000 claims description 4
- 239000004793 Polystyrene Substances 0.000 claims description 3
- 108010067390 Viral Proteins Proteins 0.000 claims description 3
- 229920002223 polystyrene Polymers 0.000 claims description 3
- 101710165845 CD81 protein Proteins 0.000 claims description 2
- 238000012360 testing method Methods 0.000 claims description 2
- 210000005260 human cell Anatomy 0.000 claims 1
- 238000002955 isolation Methods 0.000 abstract description 4
- 238000000605 extraction Methods 0.000 abstract description 3
- 239000008188 pellet Substances 0.000 description 19
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 16
- 238000001262 western blot Methods 0.000 description 11
- 210000004369 blood Anatomy 0.000 description 8
- 239000008280 blood Substances 0.000 description 8
- 241000700605 Viruses Species 0.000 description 7
- 238000005119 centrifugation Methods 0.000 description 7
- 238000000746 purification Methods 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 5
- 102000043131 MHC class II family Human genes 0.000 description 4
- 108091054438 MHC class II family Proteins 0.000 description 4
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- 210000000612 antigen-presenting cell Anatomy 0.000 description 3
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- 230000027455 binding Effects 0.000 description 3
- 238000003757 reverse transcription PCR Methods 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- 102100027221 CD81 antigen Human genes 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 101710091045 Envelope protein Proteins 0.000 description 2
- 101000914469 Homo sapiens CD82 antigen Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 101710188315 Protein X Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 102100021696 Syncytin-1 Human genes 0.000 description 2
- 108020000999 Viral RNA Proteins 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
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- 238000000338 in vitro Methods 0.000 description 2
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- 208000015181 infectious disease Diseases 0.000 description 2
- 238000007885 magnetic separation Methods 0.000 description 2
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- 238000011160 research Methods 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- 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 1
- 241000894006 Bacteria Species 0.000 description 1
- 102100027217 CD82 antigen Human genes 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
- 241000196324 Embryophyta Species 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 206010019786 Hepatitis non-A non-B Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 108700031126 Tetraspanins Proteins 0.000 description 1
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 210000001723 extracellular space Anatomy 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 208000010710 hepatitis C virus infection Diseases 0.000 description 1
- 102000055772 human CD81 Human genes 0.000 description 1
- 102000045407 human CD82 Human genes 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 210000004779 membrane envelope Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 210000002741 palatine tonsil Anatomy 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24251—Methods of production or purification of viral material
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Steroid Compounds (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB9927320.3A GB9927320D0 (en) | 1999-11-18 | 1999-11-18 | Exosome separation |
| GB9927320.3 | 1999-11-18 | ||
| PCT/IB2000/001801 WO2001036601A1 (en) | 1999-11-18 | 2000-11-20 | Method for the preparation of purified hcv rna by exosome separation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2392075A1 CA2392075A1 (en) | 2001-05-25 |
| CA2392075C true CA2392075C (en) | 2011-01-04 |
Family
ID=10864765
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2392075A Expired - Lifetime CA2392075C (en) | 1999-11-18 | 2000-11-20 | Method for the preparation of purified hcv rna by exosome separation |
Country Status (13)
| Country | Link |
|---|---|
| US (2) | US7198923B1 (enExample) |
| EP (1) | EP1230347B1 (enExample) |
| JP (1) | JP4675017B2 (enExample) |
| AT (1) | ATE373084T1 (enExample) |
| AU (1) | AU1721801A (enExample) |
| CA (1) | CA2392075C (enExample) |
| CY (1) | CY1107521T1 (enExample) |
| DE (1) | DE60036403T2 (enExample) |
| DK (1) | DK1230347T3 (enExample) |
| ES (1) | ES2292489T3 (enExample) |
| GB (1) | GB9927320D0 (enExample) |
| PT (1) | PT1230347E (enExample) |
| WO (1) | WO2001036601A1 (enExample) |
Families Citing this family (89)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9927320D0 (en) * | 1999-11-18 | 2000-01-12 | Chiron Spa | Exosome separation |
| EP1766053A4 (en) * | 2004-06-02 | 2007-12-12 | Sourcepharm Inc | MICROVESICLES CONTAINING RNA AND METHODS |
| US8021847B2 (en) | 2004-06-02 | 2011-09-20 | Proxy Life Science Holdings, Inc. | Microvesicle-based compositions and methods |
| ES2736726T3 (es) * | 2006-03-09 | 2020-01-07 | Aethlon Medical Inc | Eliminación extracorpórea de partículas microvesiculares |
| US9085778B2 (en) * | 2006-05-03 | 2015-07-21 | VL27, Inc. | Exosome transfer of nucleic acids to cells |
| US8617806B2 (en) * | 2008-01-25 | 2013-12-31 | Hansabiomed Ou | Method to measure and characterize microvesicles in the human body fluids |
| EP2604704B1 (en) * | 2008-02-01 | 2018-10-03 | The General Hospital Corporation | Use of microvesicles in diagnosis and prognosis of brain tumor |
| US20110195426A1 (en) | 2009-07-16 | 2011-08-11 | The General Hospital Corporation | Nucleic acids analysis |
| US11266736B2 (en) | 2008-04-17 | 2022-03-08 | Vin De Bona Trading Company Pte Ltd | Method of painting micro vesicles |
| EP2310529A2 (en) * | 2008-06-20 | 2011-04-20 | Proxy Life Science Holdings, Inc. | Microvesicle-based compositions and methods |
| US10545149B2 (en) * | 2008-10-06 | 2020-01-28 | Morehouse School Of Medicine | Detection of HIV-related proteins in urine |
| US9487837B2 (en) | 2008-10-06 | 2016-11-08 | Morehouse School Of Medicine | Exosome-mediated diagnosis of hepatitis virus infections and diseases |
| US7888035B2 (en) | 2008-10-30 | 2011-02-15 | Caris Mpi, Inc. | Methods for assessing RNA patterns |
| EP3181705A1 (en) * | 2008-11-12 | 2017-06-21 | Caris Life Sciences Switzerland Holdings GmbH | Methods and systems of using exosomes for determining phenotypes |
| ES2927225T3 (es) | 2009-04-17 | 2022-11-03 | Univ Oxford Innovation Ltd | Composición para entrega de material genético |
| BRPI0900815A2 (pt) * | 2009-04-23 | 2010-12-28 | Sociedade Benef Israelita Bras Hospital Albert Einstein | método para isolamento de exossomos a partir de soluções biológicas utilizando nanopartìculas de óxido de ferro |
| EP2475989A4 (en) | 2009-09-09 | 2013-02-27 | Gen Hospital Corp | USE OF MICROVESICLES IN THE ANALYSIS OF KRAS MUTATIONS |
| EP3461912B1 (en) | 2009-09-09 | 2022-07-13 | The General Hospital Corporation | Use of microvesicles in analyzing nucleic acid profiles |
| JPWO2011049059A1 (ja) * | 2009-10-22 | 2013-03-14 | 国立大学法人岐阜大学 | Ｒｎａ内包キャリア組成物 |
| EP2507393A4 (en) | 2009-11-30 | 2013-05-01 | Caris Life Sciences Luxembourg Holdings | METHODS AND SYSTEMS FOR ISOLATING, STORING AND ANALYZING VESICLES |
| CN103237901B (zh) | 2010-03-01 | 2016-08-03 | 卡里斯生命科学瑞士控股有限责任公司 | 用于治疗诊断的生物标志物 |
| CA2795776A1 (en) | 2010-04-06 | 2011-10-13 | Caris Life Sciences Luxembourg Holdings, S.A.R.L. | Circulating biomarkers for disease |
| WO2011156734A2 (en) * | 2010-06-11 | 2011-12-15 | Hitachi Chemical Co., Ltd. | Method of characterizing vascular diseases |
| JP5755326B2 (ja) | 2010-06-11 | 2015-07-29 | 日立化成株式会社 | 腎機能の特徴を決定する方法 |
| WO2012006476A2 (en) | 2010-07-07 | 2012-01-12 | Aethlon Medical, Inc. | Methods and compositions for quantifying exosomes |
| US20140045915A1 (en) | 2010-08-31 | 2014-02-13 | The General Hospital Corporation | Cancer-related biological materials in microvesicles |
| EP2638057B1 (en) | 2010-11-10 | 2019-03-06 | Exosome Diagnostics, Inc. | Method for isolation of nucleic acid containing particles and extraction of nucleic acids therefrom |
| EP3569607A1 (en) * | 2011-05-11 | 2019-11-20 | Exosome Diagnostics, Inc. | Nucleic acid extraction from heterogeneous biological materials |
| WO2012170037A1 (en) | 2011-06-10 | 2012-12-13 | Hitachi Chemical Co., Ltd. | Vesicle capturing devices and methods for using same |
| CA2839896A1 (en) | 2011-06-21 | 2012-12-27 | Alnylam Pharmaceuticals, Inc. | Assays and methods for determining activity of a therapeutic agent in a subject |
| EP2748335B1 (en) | 2011-08-22 | 2018-10-03 | Exosome Diagnostics, Inc. | Urine biomarkers |
| EP2758774A4 (en) * | 2011-09-22 | 2015-04-29 | Univ Los Andes | METHOD FOR MONITORING, DIAGNOSIS AND / OR FORECASTING ACUTE CHILDIER INJURY IN AN EARLY STAGE |
| US10174361B2 (en) | 2011-11-10 | 2019-01-08 | Exosome Diagnostics, Inc. | Cerebrospinal fluid assay |
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| CN104995311A (zh) | 2012-08-30 | 2015-10-21 | 外来体诊断公司 | 用于核酸测定的对照 |
| CN105025878A (zh) | 2012-10-03 | 2015-11-04 | 外来体诊断公司 | 微泡在医学疾病和病况的诊断、预后和治疗中的用途 |
| DK2906717T3 (en) | 2012-10-11 | 2019-04-01 | Vin De Bona Trading Company Pte Ltd | PROCEDURE FOR PAINTING MICROVESICS |
| US10942184B2 (en) | 2012-10-23 | 2021-03-09 | Caris Science, Inc. | Aptamers and uses thereof |
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| CN105209881B (zh) | 2013-03-13 | 2021-01-22 | 迈阿密大学 | 从细胞培养上清液和生物流体分离并纯化微泡的方法 |
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| KR0185373B1 (ko) * | 1989-03-17 | 1999-05-01 | 로버트 피. 블랙버언 | Hcv 폴리단백질에서 유래되는 hcv 아미노산 서열 부분을 포함하는 폴리펩티드 및 그 사용 |
| US5372928A (en) * | 1989-09-15 | 1994-12-13 | Chiron Corporation | Hepatitis C virus isolates |
| WO1991015574A1 (en) | 1990-04-03 | 1991-10-17 | Southwest Foundation For Biomedical Research | Purified hcv and hcv proteins and peptides |
| GB9416671D0 (en) | 1994-08-17 | 1994-10-12 | Biocine Spa | Assay |
| EP0841945B1 (en) * | 1995-08-03 | 2006-03-08 | Rijksuniversiteit te Leiden | Cell derived antigen presenting vesicles |
| GB9517926D0 (en) | 1995-09-01 | 1995-11-01 | Biocine Spa | Binding protein |
| FR2766205B1 (fr) * | 1997-07-16 | 2002-08-30 | Inst Nat Sante Rech Med | Nouveau procede de sensibilisation de cellules presentatrices d'antigene et nouveaux moyens pour la mise en oeuvre du procede |
| DE69841322D1 (de) * | 1997-10-06 | 2010-01-07 | Novartis Vaccines & Diagnostic | Hepatitis c rezeptorprotein cd81 |
| DE69836758T2 (de) * | 1997-11-06 | 2007-10-11 | Innogenetics N.V. | Apo b, annexin v und tubulin: medizinische und diagnostische verwendung und verwendung zur purifikation von hcv |
| GB9927320D0 (en) * | 1999-11-18 | 2000-01-12 | Chiron Spa | Exosome separation |
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- 2000-11-20 EP EP00979837A patent/EP1230347B1/en not_active Expired - Lifetime
- 2000-11-20 CA CA2392075A patent/CA2392075C/en not_active Expired - Lifetime
- 2000-11-20 WO PCT/IB2000/001801 patent/WO2001036601A1/en not_active Ceased
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| DE60036403T2 (de) | 2008-06-12 |
| US7807438B2 (en) | 2010-10-05 |
| JP4675017B2 (ja) | 2011-04-20 |
| DE60036403D1 (de) | 2007-10-25 |
| DK1230347T3 (da) | 2007-11-05 |
| JP2003514523A (ja) | 2003-04-22 |
| ATE373084T1 (de) | 2007-09-15 |
| US7198923B1 (en) | 2007-04-03 |
| GB9927320D0 (en) | 2000-01-12 |
| EP1230347A1 (en) | 2002-08-14 |
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| CY1107521T1 (el) | 2013-03-13 |
| ES2292489T3 (es) | 2008-03-16 |
| CA2392075A1 (en) | 2001-05-25 |
| PT1230347E (pt) | 2007-12-20 |
| US20070254351A1 (en) | 2007-11-01 |
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