BR112019007831A2 - ensaios in vitro e baseados em células para medição da atividade de neurotoxinas botulínicas - Google Patents
ensaios in vitro e baseados em células para medição da atividade de neurotoxinas botulínicasInfo
- Publication number
- BR112019007831A2 BR112019007831A2 BR112019007831A BR112019007831A BR112019007831A2 BR 112019007831 A2 BR112019007831 A2 BR 112019007831A2 BR 112019007831 A BR112019007831 A BR 112019007831A BR 112019007831 A BR112019007831 A BR 112019007831A BR 112019007831 A2 BR112019007831 A2 BR 112019007831A2
- Authority
- BR
- Brazil
- Prior art keywords
- neurotoxin
- polypeptides
- disclosed
- activity
- vitro
- Prior art date
Links
- 108030001720 Bontoxilysin Proteins 0.000 title abstract 2
- 231100001103 botulinum neurotoxin Toxicity 0.000 title abstract 2
- 230000007900 neurotoxin activity Effects 0.000 title abstract 2
- 238000000423 cell based assay Methods 0.000 title 1
- 238000000099 in vitro assay Methods 0.000 title 1
- 239000002581 neurotoxin Substances 0.000 abstract 4
- 231100000618 neurotoxin Toxicity 0.000 abstract 4
- 229920001184 polypeptide Polymers 0.000 abstract 4
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 4
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 4
- 101710138657 Neurotoxin Proteins 0.000 abstract 2
- 238000003776 cleavage reaction Methods 0.000 abstract 2
- 239000003446 ligand Substances 0.000 abstract 2
- 230000007017 scission Effects 0.000 abstract 2
- 101100272852 Clostridium botulinum (strain Langeland / NCTC 10281 / Type F) F gene Proteins 0.000 abstract 1
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract 1
- 108030001722 Tentoxilysin Proteins 0.000 abstract 1
- 210000004900 c-terminal fragment Anatomy 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 abstract 1
- 238000012512 characterization method Methods 0.000 abstract 1
- 230000007423 decrease Effects 0.000 abstract 1
- 238000001514 detection method Methods 0.000 abstract 1
- 238000000338 in vitro Methods 0.000 abstract 1
- 238000001727 in vivo Methods 0.000 abstract 1
- 238000000034 method Methods 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 210000004898 n-terminal fragment Anatomy 0.000 abstract 1
- 108020004707 nucleic acids Proteins 0.000 abstract 1
- 102000039446 nucleic acids Human genes 0.000 abstract 1
- 150000007523 nucleic acids Chemical class 0.000 abstract 1
- 230000004952 protein activity Effects 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 abstract 1
Classifications
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- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
-
- 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
- C12Q1/66—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving luciferase
-
- 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
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/24—Metalloendopeptidases (3.4.24)
- C12Y304/24069—Bontoxilysin (3.4.24.69), i.e. botulinum neurotoxin
-
- 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
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/33—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Clostridium (G)
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Toxicology (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
são divulgados neste documento meios para a detecção e caracterização de neurotoxinas, tais como neurotoxina botulínica (bont) ou neurotoxina tetânica. a presente divulgação fornece métodos para determinar a potência e a atividade de neurotoxinas in vitro e in vivo. são divulgados também polipeptídeos que compreendem fragmentos n-terminal e c-terminal de uma proteína repórter que são divididos por um ligante compreendendo um sítio de clivagem de neurotoxina. a clivagem do ligante por uma neurotoxina diminui a atividade da proteína repórter indicando, assim, a atividade da neurotoxina. são divulgadas também composições e kits que compreendem os polipeptídeos divulgados, ácidos nucleicos que compreendem sequências nucleotídicas que codificam tais polipeptídeos e células que expressam esses polipeptídeos.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201662410558P | 2016-10-20 | 2016-10-20 | |
| PCT/US2017/057411 WO2018075783A2 (en) | 2016-10-20 | 2017-10-19 | In vitro and cell based assays for measuring the activity of botulinum neurotoxins |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| BR112019007831A2 true BR112019007831A2 (pt) | 2019-10-01 |
Family
ID=60413254
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| BR112019007831A BR112019007831A2 (pt) | 2016-10-20 | 2017-10-19 | ensaios in vitro e baseados em células para medição da atividade de neurotoxinas botulínicas |
Country Status (19)
| Country | Link |
|---|---|
| US (2) | US11306347B2 (pt) |
| EP (1) | EP3529616B1 (pt) |
| JP (2) | JP7227901B2 (pt) |
| KR (1) | KR102556363B1 (pt) |
| CN (1) | CN110088624B (pt) |
| AU (1) | AU2017345560A1 (pt) |
| BR (1) | BR112019007831A2 (pt) |
| CA (1) | CA3040507A1 (pt) |
| DK (1) | DK3529616T3 (pt) |
| EA (1) | EA039761B1 (pt) |
| ES (1) | ES2963830T3 (pt) |
| FI (1) | FI3529616T3 (pt) |
| HU (1) | HUE064332T2 (pt) |
| MX (2) | MX2019004431A (pt) |
| PL (1) | PL3529616T3 (pt) |
| PT (1) | PT3529616T (pt) |
| SA (1) | SA519401607B1 (pt) |
| SG (1) | SG11201903523YA (pt) |
| WO (1) | WO2018075783A2 (pt) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2655614B1 (en) | 2010-12-22 | 2017-03-15 | President and Fellows of Harvard College | Continuous directed evolution |
| US10920208B2 (en) | 2014-10-22 | 2021-02-16 | President And Fellows Of Harvard College | Evolution of proteases |
| US11299729B2 (en) | 2015-04-17 | 2022-04-12 | President And Fellows Of Harvard College | Vector-based mutagenesis system |
| US10392674B2 (en) | 2015-07-22 | 2019-08-27 | President And Fellows Of Harvard College | Evolution of site-specific recombinases |
| WO2017015559A2 (en) | 2015-07-23 | 2017-01-26 | President And Fellows Of Harvard College | Evolution of bt toxins |
| US10612011B2 (en) | 2015-07-30 | 2020-04-07 | President And Fellows Of Harvard College | Evolution of TALENs |
| EP3458472B1 (en) * | 2016-05-16 | 2021-10-20 | President and Fellows of Harvard College | Method for purification and activation of botulinum neurotoxin |
| HUE064332T2 (hu) | 2016-10-20 | 2024-03-28 | Harvard College | In vitro és sejtalapú esszék botulinum neurotoxinok aktivitásának mérésére |
| US11447809B2 (en) | 2017-07-06 | 2022-09-20 | President And Fellows Of Harvard College | Evolution of tRNA synthetases |
| WO2019056002A1 (en) | 2017-09-18 | 2019-03-21 | President And Fellows Of Harvard College | CONTINUOUS EVOLUTION FOR STABILIZED PROTEINS |
| WO2019241649A1 (en) | 2018-06-14 | 2019-12-19 | President And Fellows Of Harvard College | Evolution of cytidine deaminases |
| US20220259269A1 (en) * | 2019-07-15 | 2022-08-18 | President And Fellows Of Harvard College | Evolved botulinum neurotoxins and uses thereof |
| WO2021195479A1 (en) * | 2020-03-26 | 2021-09-30 | Cellex, Inc. | Protease assays and their applications |
| CN114540419A (zh) * | 2022-03-04 | 2022-05-27 | 中国人民解放军军事科学院军事医学研究院 | 一种分析包膜病毒膜融合效率的三功能报告系统 |
| GB202213479D0 (en) | 2022-09-14 | 2022-10-26 | Ipsen Biopharm Ltd | Cell-free clostridial neurotoxin assays |
| GB202404021D0 (en) | 2024-03-20 | 2024-05-01 | Ipsen Biopharm Ltd | Cell-based neurotoxin assay |
Family Cites Families (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4737462A (en) | 1982-10-19 | 1988-04-12 | Cetus Corporation | Structural genes, plasmids and transformed cells for producing cysteine depleted muteins of interferon-β |
| US4518584A (en) | 1983-04-15 | 1985-05-21 | Cetus Corporation | Human recombinant interleukin-2 muteins |
| US4950588A (en) | 1985-07-10 | 1990-08-21 | Molecular Diagnostics, Inc. | Prolonged enhanced chemiluminescence |
| DE3537877A1 (de) | 1985-10-24 | 1987-04-30 | Geiger Reinhard | Luciferin-derivate und immunoassays unter einsatz derartiger luciferin-derivate |
| US5004565A (en) | 1986-07-17 | 1991-04-02 | The Board Of Governors Of Wayne State University | Method and compositions providing enhanced chemiluminescence from 1,2-dioxetanes |
| US5374534A (en) | 1990-07-19 | 1994-12-20 | Charm Sciences, Inc. | Method of preparing D-luciferin derivatives |
| DE4210759A1 (de) | 1992-04-01 | 1993-10-07 | Boehringer Mannheim Gmbh | Substituierte Thiazolin-Dioxetan-Substrate, Verfahren zur Herstellung und Verwendung |
| AU695623B2 (en) | 1994-05-31 | 1998-08-20 | Allergan, Inc. | Modification of clostridial toxins for use as transport proteins |
| GB9508204D0 (en) | 1995-04-21 | 1995-06-07 | Speywood Lab Ltd | A novel agent able to modify peripheral afferent function |
| GB9617671D0 (en) | 1996-08-23 | 1996-10-02 | Microbiological Res Authority | Recombinant toxin fragments |
| GB9721189D0 (en) | 1997-10-08 | 1997-12-03 | Speywood Lab The Limited | Analgesic conjugates |
| CA2324648C (en) | 1998-03-27 | 2013-02-26 | Prolume, Ltd. | Luciferases, fluorescent proteins, nucleic acids encoding the luciferases and fluorescent proteins and the use thereof in diagnostics, high throughput screening and novelty items |
| US6890745B1 (en) | 2000-07-19 | 2005-05-10 | Chemicon International, Inc. | Protease specific cleavable luciferases and methods of use thereof |
| EP2308978B8 (en) * | 2003-10-10 | 2015-04-08 | Promega Corporation | Luciferase biosensor |
| US8753831B2 (en) | 2007-06-05 | 2014-06-17 | City Of Hope | Methods for detection of botulinum neurotoxin |
| DK2456864T3 (en) | 2009-05-01 | 2015-12-21 | Promega Corp | SYNTHETIC OPLOPHORUS luciferases WITH IMPROVED LIGHT MISSION |
| US20140287433A1 (en) * | 2011-07-19 | 2014-09-25 | ETH Zürich | Means and methods for determining clostridial neurotoxins |
| AU2013229472A1 (en) | 2012-03-07 | 2014-08-07 | Merz Pharma Gmbh & Co. Kgaa | Means and methods for determining neurotoxin activity based on a modified luciferase |
| WO2014060373A1 (en) * | 2012-10-16 | 2014-04-24 | Merz Pharma Gmbh & Co. Kgaa | Cellular test systems for the determination of the biological activities of neurotoxin polypeptides |
| CN104837862A (zh) * | 2012-11-21 | 2015-08-12 | 莫茨制药有限及两合公司 | 测定肉毒杆菌神经毒素的生物活性的手段和方法 |
| ES2926463T3 (es) | 2013-03-15 | 2022-10-26 | Promega Corp | Activación de la bioluminiscencia mediante complementación estructural |
| CA2913686C (en) * | 2013-06-28 | 2021-08-03 | Merz Pharma Gmbh & Co. Kgaa | Means and methods for the determination of the biological activity of neurotoxin polypeptides in cells |
| EP3274364B1 (en) * | 2015-03-26 | 2021-08-04 | President and Fellows of Harvard College | Engineered botulinum neurotoxin |
| HUE064332T2 (hu) | 2016-10-20 | 2024-03-28 | Harvard College | In vitro és sejtalapú esszék botulinum neurotoxinok aktivitásának mérésére |
| AU2019398214C1 (en) * | 2018-12-11 | 2024-03-28 | Q32 Bio Inc. | Fusion protein constructs for complement associated disease |
-
2017
- 2017-10-19 HU HUE17801528A patent/HUE064332T2/hu unknown
- 2017-10-19 EP EP17801528.5A patent/EP3529616B1/en active Active
- 2017-10-19 KR KR1020197013429A patent/KR102556363B1/ko active IP Right Grant
- 2017-10-19 AU AU2017345560A patent/AU2017345560A1/en active Pending
- 2017-10-19 SG SG11201903523YA patent/SG11201903523YA/en unknown
- 2017-10-19 PT PT178015285T patent/PT3529616T/pt unknown
- 2017-10-19 JP JP2019521130A patent/JP7227901B2/ja active Active
- 2017-10-19 CN CN201780077166.5A patent/CN110088624B/zh active Active
- 2017-10-19 CA CA3040507A patent/CA3040507A1/en active Pending
- 2017-10-19 MX MX2019004431A patent/MX2019004431A/es unknown
- 2017-10-19 EA EA201990929A patent/EA039761B1/ru unknown
- 2017-10-19 FI FIEP17801528.5T patent/FI3529616T3/fi active
- 2017-10-19 ES ES17801528T patent/ES2963830T3/es active Active
- 2017-10-19 US US16/343,513 patent/US11306347B2/en active Active
- 2017-10-19 BR BR112019007831A patent/BR112019007831A2/pt unknown
- 2017-10-19 DK DK17801528.5T patent/DK3529616T3/da active
- 2017-10-19 WO PCT/US2017/057411 patent/WO2018075783A2/en unknown
- 2017-10-19 PL PL17801528.5T patent/PL3529616T3/pl unknown
-
2019
- 2019-04-15 MX MX2023004733A patent/MX2023004733A/es unknown
- 2019-04-18 SA SA519401607A patent/SA519401607B1/ar unknown
-
2022
- 2022-03-10 US US17/691,848 patent/US11788069B2/en active Active
- 2022-10-21 JP JP2022169039A patent/JP2023011700A/ja not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| WO2018075783A2 (en) | 2018-04-26 |
| US11788069B2 (en) | 2023-10-17 |
| EA039761B1 (ru) | 2022-03-10 |
| MX2023004733A (es) | 2023-05-10 |
| US11306347B2 (en) | 2022-04-19 |
| SA519401607B1 (ar) | 2022-08-04 |
| HUE064332T2 (hu) | 2024-03-28 |
| DK3529616T3 (da) | 2023-12-11 |
| SG11201903523YA (en) | 2019-05-30 |
| CA3040507A1 (en) | 2018-04-26 |
| KR20190068582A (ko) | 2019-06-18 |
| JP7227901B2 (ja) | 2023-02-22 |
| PT3529616T (pt) | 2023-12-06 |
| MX2019004431A (es) | 2019-08-14 |
| EP3529616A2 (en) | 2019-08-28 |
| JP2019532651A (ja) | 2019-11-14 |
| FI3529616T3 (fi) | 2023-12-04 |
| AU2017345560A1 (en) | 2019-05-02 |
| US20220356508A1 (en) | 2022-11-10 |
| PL3529616T3 (pl) | 2024-03-18 |
| EP3529616B1 (en) | 2023-09-06 |
| KR102556363B1 (ko) | 2023-07-18 |
| ES2963830T3 (es) | 2024-04-02 |
| US20190276873A1 (en) | 2019-09-12 |
| WO2018075783A3 (en) | 2018-06-21 |
| JP2023011700A (ja) | 2023-01-24 |
| EA201990929A1 (ru) | 2019-10-31 |
| CN110088624B (zh) | 2024-01-16 |
| CN110088624A (zh) | 2019-08-02 |
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