AU2017345560C1 - 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 neurotoxins Download PDF

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AU2017345560C1
AU2017345560C1 AU2017345560A AU2017345560A AU2017345560C1 AU 2017345560 C1 AU2017345560 C1 AU 2017345560C1 AU 2017345560 A AU2017345560 A AU 2017345560A AU 2017345560 A AU2017345560 A AU 2017345560A AU 2017345560 C1 AU2017345560 C1 AU 2017345560C1
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bont
ile
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AU2017345560A1 (en
AU2017345560B2 (en
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Min Dong
Feifan YU
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Harvard University
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Harvard University
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004Oxidoreductases (1.)
    • C12N9/0069Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/24Hydrolases (3) acting on glycosyl compounds (3.2)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/48Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y304/00Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/24Metalloendopeptidases (3.4.24)
    • C12Y304/24069Bontoxilysin (3.4.24.69), i.e. botulinum neurotoxin
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/66Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving luciferase
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/33Assays involving biological materials from specific organisms or of a specific nature from bacteria from Clostridium (G)
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/542Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching

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  • 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)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Toxicology (AREA)
  • Biophysics (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (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)
AU2017345560A 2016-10-20 2017-10-19 In vitro and cell based assays for measuring the activity of botulinum neurotoxins Active AU2017345560C1 (en)

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

Publications (3)

Publication Number Publication Date
AU2017345560A1 AU2017345560A1 (en) 2019-05-02
AU2017345560B2 AU2017345560B2 (en) 2024-12-12
AU2017345560C1 true AU2017345560C1 (en) 2025-04-17

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AU2017345560A Active AU2017345560C1 (en) 2016-10-20 2017-10-19 In vitro and cell based assays for measuring the activity of botulinum neurotoxins

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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)

* Cited by examiner, † Cited by third party
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

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005038029A2 (en) * 2003-10-10 2005-04-28 Promega Corporation Luciferase biosensor

Family Cites Families (24)

* Cited by examiner, † Cited by third party
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
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

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005038029A2 (en) * 2003-10-10 2005-04-28 Promega Corporation Luciferase biosensor

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Publication number Publication date
DK3529616T5 (da) 2024-09-02
PL3529616T3 (pl) 2024-03-18
KR102556363B1 (ko) 2023-07-18
UA129533C2 (uk) 2025-05-28
US20190276873A1 (en) 2019-09-12
EA039761B1 (ru) 2022-03-10
KR20190068582A (ko) 2019-06-18
EP3529616B1 (en) 2023-09-06
CA3040507A1 (en) 2018-04-26
SG11201903523YA (en) 2019-05-30
AU2017345560A1 (en) 2019-05-02
US11306347B2 (en) 2022-04-19
US11788069B2 (en) 2023-10-17
PT3529616T (pt) 2023-12-06
JP2019532651A (ja) 2019-11-14
WO2018075783A3 (en) 2018-06-21
FI3529616T3 (fi) 2023-12-04
ZA201902654B (en) 2024-08-28
AU2017345560B2 (en) 2024-12-12
EP3529616A2 (en) 2019-08-28
MX2023004733A (es) 2023-05-10
DK3529616T3 (da) 2023-12-11
JP2023011700A (ja) 2023-01-24
CN110088624B (zh) 2024-01-16
HUE064332T2 (hu) 2024-03-28
SA519401607B1 (ar) 2022-08-04
ES2963830T3 (es) 2024-04-02
US20220356508A1 (en) 2022-11-10
BR112019007831A2 (pt) 2019-10-01
WO2018075783A2 (en) 2018-04-26
CN110088624A (zh) 2019-08-02
JP7227901B2 (ja) 2023-02-22
MX2019004431A (es) 2019-08-14
EA201990929A1 (ru) 2019-10-31

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