SG11201903523YA - In vitro and cell based assays for measuring the activity of botulinum neurotoxins - Google Patents
In vitro and cell based assays for measuring the activity of botulinum neurotoxinsInfo
- Publication number
- SG11201903523YA SG11201903523YA SG11201903523YA SG11201903523YA SG11201903523YA SG 11201903523Y A SG11201903523Y A SG 11201903523YA SG 11201903523Y A SG11201903523Y A SG 11201903523YA SG 11201903523Y A SG11201903523Y A SG 11201903523YA SG 11201903523Y A SG11201903523Y A SG 11201903523YA
- Authority
- SG
- Singapore
- Prior art keywords
- international
- activity
- neurotoxin
- polypeptides
- rule
- Prior art date
Links
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
-
- 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
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- 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
-
- 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
INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property 111111101111 0 1110101011111 011101 0 01111H111001H1111111011111111111110111111 Organization International Bureau (10) International Publication Number (43) International Publication Date .....0\"\" WO 2018/075783 A2 26 April 2018 (26.04.2018) WIP0 I PCT (51) International Patent Classification: Declarations under Rule 4.17: G01N 33/542 (2006.01) Cl 2N 9/02 (2006.01) C12Q 1/66 (2006.01) — as to applicant's entitlement to apply for and be granted a patent (Rule 4.17(H)) (21) International Application Number: PCT/US2017/057411 Published: — without international search report and to be republished (22) International Filing Date: upon receipt of that report (Rule 48.2(g)) 19 October 2017 (19.10.2017) — with sequence listing part of description (Rule 5.2(a)) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/410,558 20 October 2016 (20.10.2016) US (71) Applicant: PRESIDENT AND FELLOWS OF HAR- VARD COLLEGE [US/US]; 17 Quincy Street, Cam- bridge, Massachusetts 02138 (US). (72) Inventors: DONG, Min; 12 Croft Lane, Weatogue, Con- necticut 06089 (US). YU, Feifan; 98 Dundee Court, Ab- erdeen, New Jersey 07747 (US). Agent: JARRELL, Brenda Herschbach et al.; Choate, Hall & Stewart LLP, Two International Place, Boston, (74) _ _ Massachusetts 02110 (US). = Designated States (unless otherwise indicated, for every (81) = kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, _ DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, = KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, _ MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, = = = TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. _ Designated States (unless otherwise indicated, for every — (84) _ _ kind of regional protection available): ARIPO (BW, GH, = GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, = UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, = TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, = EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, — MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). = ei M QC (54) Title: IN VITRO AND CELL BASED ASSAYS FOR MEASURING THE ACTIVITY OF BOTULINUM NEUROTOXINS IN Ir) (57) : Disclosed herein are means for the detection and characterization of neurotoxins such as botulinum neurotoxin (BoNT) I r ---- or tetanus neurotoxin. The present disclosure provides methods for determining potency and activity of neurotoxins in vitro and in vivo. 0 —. Also disclosed are polypeptides comprising N- and C- terminal fragments of a reporter protein that are split by a linker comprising GO a 1-1 neurotoxin cleavage site. Cleavage of the linker by a neurotoxin decreases reporter protein activity, thereby indicating activity of © the neurotoxin. Compositions and kits comprising the disclosed polypeptides, nucleic acids comprising nucleotide sequences encoding f'1 such polypeptides, and cells expressing such polypeptides are also disclosed. 0
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 |
|---|---|
| SG11201903523YA true SG11201903523YA (en) | 2019-05-30 |
Family
ID=60413254
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SG11201903523YA SG11201903523YA (en) | 2016-10-20 | 2017-10-19 | In vitro and cell based assays for measuring the activity of botulinum neurotoxins |
Country Status (20)
| Country | Link |
|---|---|
| US (2) | US11306347B2 (en) |
| EP (1) | EP3529616B1 (en) |
| JP (2) | JP7227901B2 (en) |
| KR (1) | KR102556363B1 (en) |
| CN (1) | CN110088624B (en) |
| AU (1) | AU2017345560A1 (en) |
| BR (1) | BR112019007831A2 (en) |
| CA (1) | CA3040507A1 (en) |
| DK (1) | DK3529616T5 (en) |
| EA (1) | EA039761B1 (en) |
| ES (1) | ES2963830T3 (en) |
| FI (1) | FI3529616T3 (en) |
| HU (1) | HUE064332T2 (en) |
| MX (2) | MX2019004431A (en) |
| PL (1) | PL3529616T3 (en) |
| PT (1) | PT3529616T (en) |
| SA (1) | SA519401607B1 (en) |
| SG (1) | SG11201903523YA (en) |
| WO (1) | WO2018075783A2 (en) |
| ZA (1) | ZA201902654B (en) |
Families Citing this family (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3202903B1 (en) | 2010-12-22 | 2020-02-12 | 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 |
| WO2016168631A1 (en) | 2015-04-17 | 2016-10-20 | 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 |
| US11524983B2 (en) | 2015-07-23 | 2022-12-13 | 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 |
| IL294014B2 (en) | 2015-10-23 | 2024-07-01 | Harvard College | Nucleobase editors and uses thereof |
| MX2018014066A (en) * | 2016-05-16 | 2019-04-04 | Harvard College | Method for purification and activation of botulinum neurotoxin. |
| PL3529616T3 (en) | 2016-10-20 | 2024-03-18 | President And Fellows Of Harvard College | In vitro and cell based assays for measuring the activity of botulinum neurotoxins |
| US11447809B2 (en) | 2017-07-06 | 2022-09-20 | President And Fellows Of Harvard College | Evolution of tRNA synthetases |
| WO2019040935A1 (en) | 2017-08-25 | 2019-02-28 | 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 |
| WO2021011579A1 (en) * | 2019-07-15 | 2021-01-21 | President And Fellows Of Harvard College | Evolved botulinum neurotoxins and uses thereof |
| US20210301319A1 (en) * | 2020-03-26 | 2021-09-30 | Cellex, Inc. | Protease assays and their applications |
| CN114540419A (en) * | 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 | Three-function report system for analyzing fusion efficiency of enveloped virus membrane |
| 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 |
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| 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 (en) | 1985-10-24 | 1987-04-30 | Geiger Reinhard | LUCIFERIN DERIVATIVES AND IMMUNOASSAYS USING SUCH LUCIFERIN DERIVATIVES |
| 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 (en) | 1992-04-01 | 1993-10-07 | Boehringer Mannheim Gmbh | Substituted thiazoline dioxetane substrates, process for their preparation and use |
| JP3523879B2 (en) | 1994-05-31 | 2004-04-26 | アレルガン インコーポレイテッド | Modification of Clostridium toxin for 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 |
| DE69938293T2 (en) | 1998-03-27 | 2009-03-12 | Bruce J. Beverly Hills Bryan | LUCIFERASE, GFP FLUORESCENCE PROTEINS, CODING NUCLEIC CAUSE, AND ITS USE IN DIAGNOSIS |
| US6890745B1 (en) * | 2000-07-19 | 2005-05-10 | Chemicon International, Inc. | Protease specific cleavable luciferases and methods of use thereof |
| JP2007508014A (en) * | 2003-10-10 | 2007-04-05 | プロメガ コーポレイション | Luciferase biosensor |
| US8753831B2 (en) | 2007-06-05 | 2014-06-17 | City Of Hope | Methods for detection of botulinum neurotoxin |
| PL3409764T3 (en) | 2009-05-01 | 2020-07-13 | Promega Corporation | Synthetic oplophorus luciferases with enhanced light output |
| US20140287433A1 (en) * | 2011-07-19 | 2014-09-25 | ETH Zürich | Means and methods for determining clostridial neurotoxins |
| WO2013131991A1 (en) | 2012-03-07 | 2013-09-12 | Merz Pharma Gmbh & Co. Kgaa | Means and methods for determining neurotoxin activity based on a modified luciferase |
| KR102073111B1 (en) * | 2012-10-16 | 2020-02-04 | 메르츠 파마 게엠베하 운트 코. 카가아 | Cellular test systems for the determination of the biological activities of Neurotoxin polypeptides |
| EP2922866B1 (en) * | 2012-11-21 | 2019-01-23 | Merz Pharma GmbH & Co. KGaA | Means and methods for determination of botulinum neurotoxin biological activity |
| BR112015023394B8 (en) | 2013-03-15 | 2023-09-26 | Promega Corp | Bioluminescent system and complex |
| RU2704808C2 (en) * | 2013-06-28 | 2019-10-31 | Мерц Фарма Гмбх Энд Ко. Кгаа | Agents and methods for determining biological activity of neurotoxin polypeptides in cells |
| EP3274364B1 (en) * | 2015-03-26 | 2021-08-04 | President and Fellows of Harvard College | Engineered botulinum neurotoxin |
| PL3529616T3 (en) | 2016-10-20 | 2024-03-18 | President And Fellows Of Harvard College | In vitro and cell based assays for measuring the activity of botulinum neurotoxins |
| SG11202106073QA (en) * | 2018-12-11 | 2021-07-29 | Q32 Bio Inc | Fusion protein constructs for complement associated disease |
-
2017
- 2017-10-19 PL PL17801528.5T patent/PL3529616T3/en unknown
- 2017-10-19 EP EP17801528.5A patent/EP3529616B1/en active Active
- 2017-10-19 CA CA3040507A patent/CA3040507A1/en active Pending
- 2017-10-19 DK DK17801528.5T patent/DK3529616T5/en active
- 2017-10-19 CN CN201780077166.5A patent/CN110088624B/en active Active
- 2017-10-19 SG SG11201903523YA patent/SG11201903523YA/en unknown
- 2017-10-19 PT PT178015285T patent/PT3529616T/en unknown
- 2017-10-19 MX MX2019004431A patent/MX2019004431A/en unknown
- 2017-10-19 FI FIEP17801528.5T patent/FI3529616T3/en active
- 2017-10-19 BR BR112019007831A patent/BR112019007831A2/en unknown
- 2017-10-19 KR KR1020197013429A patent/KR102556363B1/en active IP Right Grant
- 2017-10-19 EA EA201990929A patent/EA039761B1/en unknown
- 2017-10-19 US US16/343,513 patent/US11306347B2/en active Active
- 2017-10-19 ES ES17801528T patent/ES2963830T3/en active Active
- 2017-10-19 AU AU2017345560A patent/AU2017345560A1/en active Pending
- 2017-10-19 HU HUE17801528A patent/HUE064332T2/en unknown
- 2017-10-19 JP JP2019521130A patent/JP7227901B2/en active Active
- 2017-10-19 WO PCT/US2017/057411 patent/WO2018075783A2/en unknown
-
2019
- 2019-04-15 MX MX2023004733A patent/MX2023004733A/en unknown
- 2019-04-18 SA SA519401607A patent/SA519401607B1/en unknown
- 2019-04-26 ZA ZA2019/02654A patent/ZA201902654B/en unknown
-
2022
- 2022-03-10 US US17/691,848 patent/US11788069B2/en active Active
- 2022-10-21 JP JP2022169039A patent/JP2023011700A/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| HUE064332T2 (en) | 2024-03-28 |
| KR20190068582A (en) | 2019-06-18 |
| US20220356508A1 (en) | 2022-11-10 |
| EA201990929A1 (en) | 2019-10-31 |
| DK3529616T3 (en) | 2023-12-11 |
| CA3040507A1 (en) | 2018-04-26 |
| JP7227901B2 (en) | 2023-02-22 |
| EA039761B1 (en) | 2022-03-10 |
| SA519401607B1 (en) | 2022-08-04 |
| JP2023011700A (en) | 2023-01-24 |
| PT3529616T (en) | 2023-12-06 |
| US11306347B2 (en) | 2022-04-19 |
| US20190276873A1 (en) | 2019-09-12 |
| JP2019532651A (en) | 2019-11-14 |
| WO2018075783A2 (en) | 2018-04-26 |
| MX2023004733A (en) | 2023-05-10 |
| AU2017345560A1 (en) | 2019-05-02 |
| KR102556363B1 (en) | 2023-07-18 |
| BR112019007831A2 (en) | 2019-10-01 |
| MX2019004431A (en) | 2019-08-14 |
| CN110088624A (en) | 2019-08-02 |
| ES2963830T3 (en) | 2024-04-02 |
| CN110088624B (en) | 2024-01-16 |
| DK3529616T5 (en) | 2024-09-02 |
| WO2018075783A3 (en) | 2018-06-21 |
| PL3529616T3 (en) | 2024-03-18 |
| US11788069B2 (en) | 2023-10-17 |
| ZA201902654B (en) | 2024-08-28 |
| EP3529616A2 (en) | 2019-08-28 |
| FI3529616T3 (en) | 2023-12-04 |
| EP3529616B1 (en) | 2023-09-06 |
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