JP2019532651A5 - - Google Patents
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- Publication number
- JP2019532651A5 JP2019532651A5 JP2019521130A JP2019521130A JP2019532651A5 JP 2019532651 A5 JP2019532651 A5 JP 2019532651A5 JP 2019521130 A JP2019521130 A JP 2019521130A JP 2019521130 A JP2019521130 A JP 2019521130A JP 2019532651 A5 JP2019532651 A5 JP 2019532651A5
- Authority
- JP
- Japan
- Prior art keywords
- chain polypeptide
- neurotoxin
- terminal fragment
- cleavage site
- amino acids
- 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.)
- Granted
Links
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 119
- 229920001184 polypeptide Polymers 0.000 claims description 118
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 118
- 108030001720 Bontoxilysin Proteins 0.000 claims description 68
- 231100001103 botulinum neurotoxin Toxicity 0.000 claims description 68
- 150000001413 amino acids Chemical class 0.000 claims description 66
- 108060001084 Luciferase Proteins 0.000 claims description 59
- 239000005089 Luciferase Substances 0.000 claims description 57
- 238000003776 cleavage reaction Methods 0.000 claims description 55
- 230000007017 scission Effects 0.000 claims description 55
- 238000000034 method Methods 0.000 claims description 40
- 125000006850 spacer group Chemical group 0.000 claims description 39
- 101710138657 Neurotoxin Proteins 0.000 claims description 27
- 239000002581 neurotoxin Substances 0.000 claims description 27
- 231100000618 neurotoxin Toxicity 0.000 claims description 27
- 150000007523 nucleic acids Chemical class 0.000 claims description 26
- 108020004707 nucleic acids Proteins 0.000 claims description 25
- 102000039446 nucleic acids Human genes 0.000 claims description 25
- 230000000694 effects Effects 0.000 claims description 23
- 210000004898 n-terminal fragment Anatomy 0.000 claims description 19
- 210000004899 c-terminal region Anatomy 0.000 claims description 17
- 239000012634 fragment Substances 0.000 claims description 14
- 210000004900 c-terminal fragment Anatomy 0.000 claims description 13
- 239000013598 vector Substances 0.000 claims description 13
- 102000004169 proteins and genes Human genes 0.000 claims description 12
- 108090000623 proteins and genes Proteins 0.000 claims description 12
- 210000004027 cell Anatomy 0.000 claims description 11
- 210000002569 neuron Anatomy 0.000 claims description 11
- 101000584382 Homo sapiens Synaptic vesicle glycoprotein 2C Proteins 0.000 claims description 10
- 101000652315 Homo sapiens Synaptosomal-associated protein 25 Proteins 0.000 claims description 10
- 102000053886 human SNAP25 Human genes 0.000 claims description 10
- 108020001507 fusion proteins Proteins 0.000 claims description 8
- 102000037865 fusion proteins Human genes 0.000 claims description 8
- 102000000583 SNARE Proteins Human genes 0.000 claims description 7
- 108010041948 SNARE Proteins Proteins 0.000 claims description 7
- 102000056327 human SV2C Human genes 0.000 claims description 7
- 101000807882 Homo sapiens Vesicle-associated membrane protein 1 Proteins 0.000 claims description 5
- 241000242743 Renilla reniformis Species 0.000 claims description 5
- 241000607598 Vibrio Species 0.000 claims description 5
- 125000000539 amino acid group Chemical group 0.000 claims description 5
- 238000012217 deletion Methods 0.000 claims description 5
- 230000037430 deletion Effects 0.000 claims description 5
- 102000056725 human VAMP1 Human genes 0.000 claims description 5
- 239000006166 lysate Substances 0.000 claims description 5
- 238000006467 substitution reaction Methods 0.000 claims description 5
- 238000007792 addition Methods 0.000 claims description 4
- 108090000363 Bacterial Luciferases Proteins 0.000 claims description 3
- 108090000331 Firefly luciferases Proteins 0.000 claims description 3
- 108010010469 Qa-SNARE Proteins Proteins 0.000 claims description 3
- 102000005917 R-SNARE Proteins Human genes 0.000 claims description 3
- 108010005730 R-SNARE Proteins Proteins 0.000 claims description 3
- 102100030637 Synaptic vesicle glycoprotein 2C Human genes 0.000 claims description 3
- 102000004183 Synaptosomal-Associated Protein 25 Human genes 0.000 claims description 3
- 108010057722 Synaptosomal-Associated Protein 25 Proteins 0.000 claims description 3
- 102000050389 Syntaxin Human genes 0.000 claims description 3
- 108030001722 Tentoxilysin Proteins 0.000 claims description 3
- 239000002773 nucleotide Substances 0.000 claims description 3
- 125000003729 nucleotide group Chemical group 0.000 claims description 3
- 229920002704 polyhistidine Polymers 0.000 claims description 3
- ATCJTYORYKLVIA-SRXJVYAUSA-N vamp regimen Chemical compound O=C1C=C[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1.C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1.O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1.C([C@H](C[C@]1(C(=O)OC)C=2C(=CC3=C(C45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C=O)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 ATCJTYORYKLVIA-SRXJVYAUSA-N 0.000 claims description 3
- 241000607620 Aliivibrio fischeri Species 0.000 claims description 2
- 241000193155 Clostridium botulinum Species 0.000 claims description 2
- 244000225942 Viola tricolor Species 0.000 claims description 2
- 235000004031 Viola x wittrockiana Nutrition 0.000 claims description 2
- 101000874762 Homo sapiens Synaptotagmin-2 Proteins 0.000 claims 1
- 241000288906 Primates Species 0.000 claims 1
- 102100036151 Synaptotagmin-2 Human genes 0.000 claims 1
- 241000607618 Vibrio harveyi Species 0.000 claims 1
- 239000013604 expression vector Substances 0.000 description 7
- HBZBAMXERPYTFS-SECBINFHSA-N (4S)-2-(6,7-dihydro-5H-pyrrolo[3,2-f][1,3]benzothiazol-2-yl)-4,5-dihydro-1,3-thiazole-4-carboxylic acid Chemical compound OC(=O)[C@H]1CSC(=N1)c1nc2cc3CCNc3cc2s1 HBZBAMXERPYTFS-SECBINFHSA-N 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 241000254058 Photinus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 102000004603 Vesicle-Associated Membrane Protein 1 Human genes 0.000 description 1
- 108010017743 Vesicle-Associated Membrane Protein 1 Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2022169039A JP2023011700A (ja) | 2016-10-20 | 2022-10-21 | ボツリヌス神経毒素の活性を測定するためのインビトロアッセイ法及び細胞ベースのアッセイ法 |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201662410558P | 2016-10-20 | 2016-10-20 | |
| US62/410,558 | 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 |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2022169039A Division JP2023011700A (ja) | 2016-10-20 | 2022-10-21 | ボツリヌス神経毒素の活性を測定するためのインビトロアッセイ法及び細胞ベースのアッセイ法 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2019532651A JP2019532651A (ja) | 2019-11-14 |
| JP2019532651A5 true JP2019532651A5 (enExample) | 2020-11-19 |
| JP7227901B2 JP7227901B2 (ja) | 2023-02-22 |
Family
ID=60413254
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019521130A Active JP7227901B2 (ja) | 2016-10-20 | 2017-10-19 | ボツリヌス神経毒素の活性を測定するためのインビトロアッセイ法及び細胞ベースのアッセイ法 |
| JP2022169039A Withdrawn JP2023011700A (ja) | 2016-10-20 | 2022-10-21 | ボツリヌス神経毒素の活性を測定するためのインビトロアッセイ法及び細胞ベースのアッセイ法 |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2022169039A Withdrawn JP2023011700A (ja) | 2016-10-20 | 2022-10-21 | ボツリヌス神経毒素の活性を測定するためのインビトロアッセイ法及び細胞ベースのアッセイ法 |
Country Status (21)
| Country | Link |
|---|---|
| US (2) | US11306347B2 (enExample) |
| EP (1) | EP3529616B1 (enExample) |
| JP (2) | JP7227901B2 (enExample) |
| KR (1) | KR102556363B1 (enExample) |
| CN (1) | CN110088624B (enExample) |
| AU (1) | AU2017345560C1 (enExample) |
| BR (1) | BR112019007831A2 (enExample) |
| CA (1) | CA3040507A1 (enExample) |
| DK (1) | DK3529616T5 (enExample) |
| EA (1) | EA039761B1 (enExample) |
| ES (1) | ES2963830T3 (enExample) |
| FI (1) | FI3529616T3 (enExample) |
| HU (1) | HUE064332T2 (enExample) |
| MX (2) | MX2019004431A (enExample) |
| PL (1) | PL3529616T3 (enExample) |
| PT (1) | PT3529616T (enExample) |
| SA (1) | SA519401607B1 (enExample) |
| SG (1) | SG11201903523YA (enExample) |
| UA (1) | UA129533C2 (enExample) |
| WO (1) | WO2018075783A2 (enExample) |
| ZA (1) | ZA201902654B (enExample) |
Families Citing this family (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2011348204B2 (en) | 2010-12-22 | 2017-03-02 | President And Fellows Of Harvard College | Continuous directed evolution |
| WO2015134121A2 (en) | 2014-01-20 | 2015-09-11 | President And Fellows Of Harvard College | Negative selection and stringency modulation in continuous evolution systems |
| 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 |
| 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 |
| EP3365356B1 (en) | 2015-10-23 | 2023-06-28 | President and Fellows of Harvard College | Nucleobase editors and uses thereof |
| CN109641941B (zh) * | 2016-05-16 | 2023-06-16 | 哈佛大学校长及研究员协会 | 用于纯化和激活肉毒杆菌神经毒素的方法 |
| EA039761B1 (ru) | 2016-10-20 | 2022-03-10 | Президент Энд Феллоуз Оф Гарвард Колледж | In vitro и клеточные анализы измерения активности ботулинических нейротоксинов |
| WO2019010164A1 (en) | 2017-07-06 | 2019-01-10 | President And Fellows Of Harvard College | Evolution of trna synthetases |
| EP3673059A4 (en) | 2017-08-25 | 2021-09-01 | President And Fellows Of Harvard College | EVOLUTION OF BONT PEPTIDASES |
| US11624130B2 (en) | 2017-09-18 | 2023-04-11 | President And Fellows Of Harvard College | Continuous evolution for stabilized proteins |
| US11913044B2 (en) | 2018-06-14 | 2024-02-27 | 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 |
| 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 |
| CN121057740A (zh) * | 2023-04-21 | 2025-12-02 | 深圳琅技生命科技有限公司 | 一种肽以及含有该肽的发光复合物 |
| GB202404021D0 (en) | 2024-03-20 | 2024-05-01 | Ipsen Biopharm Ltd | Cell-based neurotoxin assay |
| CN119039432B (zh) * | 2024-10-31 | 2025-01-28 | 兰州生物技术开发有限公司 | 一种检测肉毒毒素活性或突触体相关蛋白的方法和试剂盒 |
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 |
| ATE388224T1 (de) | 1998-03-27 | 2008-03-15 | Prolume Ltd | Luciferase, gfp fluoreszenzproteine, kodierende nukleinsaüre und ihre verwendung in der diagnose |
| 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 |
| SG10201901453PA (en) | 2009-05-01 | 2019-03-28 | Promega Corp | Synthetic oplophorus luciferases with enhanced light output |
| 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 |
| EP2909316B1 (en) * | 2012-10-16 | 2018-09-05 | Merz Pharma GmbH & Co. KGaA | Cellular test systems for the determination of the biological activities of neurotoxin polypeptides |
| HK1210190A1 (en) * | 2012-11-21 | 2016-04-15 | Merz Pharma Gmbh & Co. Kgaa. | Means and methods for determination of botulinum neurotoxin biological activity |
| KR102675860B1 (ko) | 2013-03-15 | 2024-06-20 | 프로메가 코포레이션 | 구조적 보완에 의한 바이오발광성의 활성화 |
| MX360513B (es) * | 2013-06-28 | 2018-11-07 | Merz Pharma Gmbh & Co Kgaa | Medios y metodos para la determinacion de la actividad biologica de polipeptidos de neurotoxina en celulas. |
| ES2895853T3 (es) * | 2015-03-26 | 2022-02-22 | Harvard College | Neurotoxina botulínica modificada |
| EA039761B1 (ru) | 2016-10-20 | 2022-03-10 | Президент Энд Феллоуз Оф Гарвард Колледж | In vitro и клеточные анализы измерения активности ботулинических нейротоксинов |
| MA54468A (fr) * | 2018-12-11 | 2022-04-13 | Q32 Bio Inc | Constructions de protéines de fusion pour une maladie associée au complément |
-
2017
- 2017-10-19 EA EA201990929A patent/EA039761B1/ru unknown
- 2017-10-19 UA UAA201905231A patent/UA129533C2/uk unknown
- 2017-10-19 PT PT178015285T patent/PT3529616T/pt unknown
- 2017-10-19 EP EP17801528.5A patent/EP3529616B1/en active Active
- 2017-10-19 ES ES17801528T patent/ES2963830T3/es active Active
- 2017-10-19 PL PL17801528.5T patent/PL3529616T3/pl unknown
- 2017-10-19 HU HUE17801528A patent/HUE064332T2/hu unknown
- 2017-10-19 MX MX2019004431A patent/MX2019004431A/es unknown
- 2017-10-19 CA CA3040507A patent/CA3040507A1/en active Pending
- 2017-10-19 WO PCT/US2017/057411 patent/WO2018075783A2/en not_active Ceased
- 2017-10-19 CN CN201780077166.5A patent/CN110088624B/zh active Active
- 2017-10-19 AU AU2017345560A patent/AU2017345560C1/en active Active
- 2017-10-19 BR BR112019007831-2A patent/BR112019007831A2/pt active Search and Examination
- 2017-10-19 US US16/343,513 patent/US11306347B2/en active Active
- 2017-10-19 DK DK17801528.5T patent/DK3529616T5/da active
- 2017-10-19 JP JP2019521130A patent/JP7227901B2/ja active Active
- 2017-10-19 SG SG11201903523YA patent/SG11201903523YA/en unknown
- 2017-10-19 FI FIEP17801528.5T patent/FI3529616T3/fi active
- 2017-10-19 KR KR1020197013429A patent/KR102556363B1/ko active Active
-
2019
- 2019-04-15 MX MX2023004733A patent/MX2023004733A/es unknown
- 2019-04-18 SA SA519401607A patent/SA519401607B1/ar 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/ja not_active Withdrawn
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