BRPI0514318A - ensaio de liberação de fluorescência com relação à atividade de protease da toxina do botulismo em gfp-snap25 - Google Patents
ensaio de liberação de fluorescência com relação à atividade de protease da toxina do botulismo em gfp-snap25Info
- Publication number
- BRPI0514318A BRPI0514318A BRPI0514318-7A BRPI0514318A BRPI0514318A BR PI0514318 A BRPI0514318 A BR PI0514318A BR PI0514318 A BRPI0514318 A BR PI0514318A BR PI0514318 A BRPI0514318 A BR PI0514318A
- Authority
- BR
- Brazil
- Prior art keywords
- snap25
- gfp
- fluorescent protein
- partner
- cleavage site
- Prior art date
Links
- 231100000765 toxin Toxicity 0.000 title abstract 3
- 239000003053 toxin Substances 0.000 title abstract 3
- 208000003508 Botulism Diseases 0.000 title abstract 2
- 108091005804 Peptidases Proteins 0.000 title abstract 2
- 239000004365 Protease Substances 0.000 title abstract 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 title abstract 2
- 238000003556 assay Methods 0.000 title 1
- 238000003776 cleavage reaction Methods 0.000 abstract 5
- 230000007017 scission Effects 0.000 abstract 5
- 108030001720 Bontoxilysin Proteins 0.000 abstract 3
- 108010043121 Green Fluorescent Proteins Proteins 0.000 abstract 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 abstract 2
- 102000004183 Synaptosomal-Associated Protein 25 Human genes 0.000 abstract 2
- 108010057722 Synaptosomal-Associated Protein 25 Proteins 0.000 abstract 2
- 108091006047 fluorescent proteins Proteins 0.000 abstract 2
- 102000034287 fluorescent proteins Human genes 0.000 abstract 2
- 239000005090 green fluorescent protein Substances 0.000 abstract 2
- 102000039446 nucleic acids Human genes 0.000 abstract 2
- 108020004707 nucleic acids Proteins 0.000 abstract 2
- 150000007523 nucleic acids Chemical class 0.000 abstract 2
- 239000002773 nucleotide Substances 0.000 abstract 2
- 125000003729 nucleotide group Chemical group 0.000 abstract 2
- 239000000758 substrate Substances 0.000 abstract 2
- 231100001102 clostridial toxin Toxicity 0.000 abstract 1
Classifications
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- 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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
-
- 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
-
- 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
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43595—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from coelenteratae, e.g. medusae
-
- 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
- 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
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
-
- 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
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- 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)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Toxicology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Food Science & Technology (AREA)
- Virology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
ENSAIO DE LIBERAçãO DE FLUORESCêNCIA COM RELAçãO à ATIVIDADE DE PROTEASE DA TOXINA DO BOTULISMO EM GFP-SNAP25. A presente invenção refere-se a uma molécula de ácido nucléico que contém uma seqüência de nucleotídeo que codifica um substrato SNAP-25 que inclui (1) uma proteína fluorescente verde; (ii) um primeiro parceiro de uma dupla de afinidade; e (iii) uma porção de SNAP-25 que inclui uma seqüência de reconhecimento BoNT/A, BoNT/C1 ou BoNT/E contendo um sítio de clivagem, em que o sitio de clivagem intervém entre a proteína fluorescente verde e o primeiro parceiro da dupla de afinidade. Também é provido aqui, neste pedido de patente, uma molécula de ácido nucléico que contêm uma seqüência de nucleotídeo que codifica um substrato de toxina alvo que inclui (1) uma proteína fluorescente, (ii) um primeiro parceiro de uma dupla de afinidade; e (iii) uma seqüência de reconhecimento da toxina do clostrídico contendo um sítio de clivagem, em que o sitio de clivagem intervém entre a proteína fluorescente e o primeiro parceiro da dupla de afinidade.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US10/917,844 US7374896B2 (en) | 2001-08-28 | 2004-08-13 | GFP-SNAP25 fluorescence release assay for botulinum neurotoxin protease activity |
| PCT/US2005/028497 WO2006020748A2 (en) | 2004-08-13 | 2005-08-10 | Gfp-snap25 fluorescence release assay for botulinum toxin protease activity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| BRPI0514318A true BRPI0514318A (pt) | 2008-06-10 |
Family
ID=35908131
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| BRPI0514318-7A BRPI0514318A (pt) | 2004-08-13 | 2005-08-10 | ensaio de liberação de fluorescência com relação à atividade de protease da toxina do botulismo em gfp-snap25 |
Country Status (7)
| Country | Link |
|---|---|
| US (5) | US7374896B2 (pt) |
| EP (1) | EP1776380A2 (pt) |
| JP (1) | JP2008509667A (pt) |
| AU (1) | AU2005272796A1 (pt) |
| BR (1) | BRPI0514318A (pt) |
| CA (1) | CA2576354A1 (pt) |
| WO (1) | WO2006020748A2 (pt) |
Families Citing this family (51)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080064054A1 (en) * | 2001-08-28 | 2008-03-13 | Ester Fernandez-Salas | Fluorescence resonance energy transfer (fret) assays for clostridial toxin activity |
| US7208285B2 (en) * | 2001-08-28 | 2007-04-24 | Allergan, Inc. | Fret protease assays for botulinum serotype A/E toxins |
| US8022172B2 (en) * | 2001-08-28 | 2011-09-20 | Allergan, Inc. | Luminescence resonance energy transfer (LRET) assays for clostridial toxin activity |
| US7332567B2 (en) * | 2001-08-28 | 2008-02-19 | Allergan, Inc. | Fret protease assays for clostridial toxins |
| US7374896B2 (en) * | 2001-08-28 | 2008-05-20 | Allergan, Inc. | GFP-SNAP25 fluorescence release assay for botulinum neurotoxin protease activity |
| CA2432676A1 (en) * | 2002-07-31 | 2004-01-31 | Robert Menard | Development of an in vivo functional assay for proteases |
| US7183066B2 (en) * | 2002-09-27 | 2007-02-27 | Allergan, Inc. | Cell-based fluorescence resonance energy transfer (FRET) assays for clostridial toxins |
| WO2004031355A2 (en) * | 2002-10-01 | 2004-04-15 | University Of Maryland | Methods for identifying inhibitors of botulinum neurotoxins |
| WO2005076785A2 (en) * | 2003-12-19 | 2005-08-25 | Wisconsin Alumni Research Foundation | Method and compositions for detecting botulinum neurotoxin |
| US7399607B2 (en) * | 2004-09-22 | 2008-07-15 | Allergan, Inc. | Fluorescence polarization assays for determining clostridial toxin activity |
| EP2293064B1 (en) | 2005-04-05 | 2012-04-04 | Allergan, Inc. | Lipophilic dye-based FRET assays for clostridial toxin activity |
| CA2605160A1 (en) * | 2005-04-05 | 2006-10-05 | Allergan, Inc. | Clostridial toxin activity assays |
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| KR101923847B1 (ko) * | 2009-03-13 | 2018-11-29 | 알러간, 인코포레이티드 | 면역 기반 재표적화된 엔도펩티다제 활성 검정 |
| CA2825370A1 (en) | 2010-12-22 | 2012-06-28 | President And Fellows Of Harvard College | Continuous directed evolution |
| EP2578695A1 (en) * | 2011-10-05 | 2013-04-10 | Albert-Ludwigs-Universität Freiburg | Assay for Clostridium botulinum Neurotoxin |
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| WO2015134121A2 (en) | 2014-01-20 | 2015-09-11 | President And Fellows Of Harvard College | Negative selection and stringency modulation in continuous evolution systems |
| ES2642916T3 (es) | 2014-06-06 | 2017-11-20 | Galit KLEINER-FISMAN | Toxina botulínica para su uso en el tratamiento de la paratonia |
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| KR102409293B1 (ko) * | 2014-11-21 | 2022-06-14 | 메르츠 파마 게엠베하 운트 코. 카가아 | 신경독소 폴리펩티드의 생물학적 활성의 측정 방법 |
| KR102514666B1 (ko) | 2014-12-19 | 2023-03-27 | 메르츠 파마 게엠베하 운트 코. 카가아 | 세포에서 BoNT/E의 생물학적 활성의 결정을 위한 수단 및 방법 |
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| US12060553B2 (en) | 2017-08-25 | 2024-08-13 | President And Fellows Of Harvard College | Evolution of BoNT peptidases |
| 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 |
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| WO2004031355A2 (en) | 2002-10-01 | 2004-04-15 | University Of Maryland | Methods for identifying inhibitors of botulinum neurotoxins |
| US20040115727A1 (en) | 2002-12-11 | 2004-06-17 | Allergan, Inc., A Corporation | Evolved clostridial toxins with altered protease specificity |
| US7172764B2 (en) * | 2003-11-17 | 2007-02-06 | Allergan, Inc. | Rescue agents for treating botulinum toxin intoxications |
| WO2005076785A2 (en) * | 2003-12-19 | 2005-08-25 | Wisconsin Alumni Research Foundation | Method and compositions for detecting botulinum neurotoxin |
| ES2374822T3 (es) * | 2004-02-24 | 2012-02-22 | Allergan, Inc. | Ensayo de selección de toxinas botul�?nicas. |
| WO2005121354A2 (fr) * | 2004-05-05 | 2005-12-22 | Pharmaleads | Substrats peptidiques reconnus par la toxine botulique de type a, bont/a et leurs utilisations |
| US7514088B2 (en) * | 2005-03-15 | 2009-04-07 | Allergan, Inc. | Multivalent Clostridial toxin derivatives and methods of their use |
| JP2006086467A (ja) * | 2004-09-17 | 2006-03-30 | Toshiba Corp | 半導体装置及びその製造方法 |
| US7399607B2 (en) * | 2004-09-22 | 2008-07-15 | Allergan, Inc. | Fluorescence polarization assays for determining clostridial toxin activity |
| EP2293064B1 (en) * | 2005-04-05 | 2012-04-04 | Allergan, Inc. | Lipophilic dye-based FRET assays for clostridial toxin activity |
| ATE518882T1 (de) * | 2005-09-19 | 2011-08-15 | Allergan Inc | Mit clostridientoxin aktivierbare clostridientoxine |
| CA2602972C (en) * | 2005-10-12 | 2013-09-24 | Allergan, Inc. | Assays of molecular or subcellular interactivity using depolarization after resonance energy transfer (daret) |
| EP2001902B1 (en) * | 2006-03-14 | 2013-03-27 | Allergan, Inc. | Modified clostridial toxins with altered targeting capabilities for clostridial toxin target cells |
| WO2009035476A1 (en) * | 2007-09-14 | 2009-03-19 | Biosentinel, Llc | Resonance energy transfer assay with cleavage sequence and spacer |
-
2004
- 2004-08-13 US US10/917,844 patent/US7374896B2/en not_active Expired - Lifetime
-
2005
- 2005-08-10 EP EP05826811A patent/EP1776380A2/en not_active Withdrawn
- 2005-08-10 JP JP2007525782A patent/JP2008509667A/ja active Pending
- 2005-08-10 CA CA002576354A patent/CA2576354A1/en not_active Abandoned
- 2005-08-10 AU AU2005272796A patent/AU2005272796A1/en not_active Abandoned
- 2005-08-10 BR BRPI0514318-7A patent/BRPI0514318A/pt not_active Application Discontinuation
- 2005-08-10 WO PCT/US2005/028497 patent/WO2006020748A2/en not_active Ceased
-
2008
- 2008-01-23 US US12/018,637 patent/US20080166739A1/en not_active Abandoned
- 2008-01-24 US US12/019,188 patent/US20080171348A1/en not_active Abandoned
- 2008-01-24 US US12/019,120 patent/US7838260B2/en not_active Expired - Lifetime
- 2008-01-24 US US12/019,070 patent/US20080176249A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| US20080176249A1 (en) | 2008-07-24 |
| US20060154314A9 (en) | 2006-07-13 |
| CA2576354A1 (en) | 2006-03-23 |
| EP1776380A2 (en) | 2007-04-25 |
| US7838260B2 (en) | 2010-11-23 |
| US20080171348A1 (en) | 2008-07-17 |
| US20050100973A1 (en) | 2005-05-12 |
| US20080166739A1 (en) | 2008-07-10 |
| JP2008509667A (ja) | 2008-04-03 |
| US20080213796A1 (en) | 2008-09-04 |
| AU2005272796A1 (en) | 2006-02-23 |
| WO2006020748A2 (en) | 2006-02-23 |
| US7374896B2 (en) | 2008-05-20 |
| WO2006020748A3 (en) | 2006-10-05 |
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