CA3040507A1 - 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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Publication number
CA3040507A1
CA3040507A1 CA3040507A CA3040507A CA3040507A1 CA 3040507 A1 CA3040507 A1 CA 3040507A1 CA 3040507 A CA3040507 A CA 3040507A CA 3040507 A CA3040507 A CA 3040507A CA 3040507 A1 CA3040507 A1 CA 3040507A1
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Prior art keywords
luciferase
terminal
amino acids
bont
single chain
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Pending
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CA3040507A
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English (en)
French (fr)
Inventor
Min Dong
Feifan YU
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Harvard University
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Harvard University
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Publication of CA3040507A1 publication Critical patent/CA3040507A1/en
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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
    • 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
    • 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
    • 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
    • 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)

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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)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Toxicology (AREA)
  • Biophysics (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (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)
CA3040507A 2016-10-20 2017-10-19 In vitro and cell based assays for measuring the activity of botulinum neurotoxins Pending CA3040507A1 (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 (1)

Publication Number Publication Date
CA3040507A1 true CA3040507A1 (en) 2018-04-26

Family

ID=60413254

Family Applications (1)

Application Number Title Priority Date Filing Date
CA3040507A Pending CA3040507A1 (en) 2016-10-20 2017-10-19 In vitro and cell based assays for measuring the activity of botulinum neurotoxins

Country Status (21)

Country Link
US (2) US11306347B2 (https=)
EP (1) EP3529616B1 (https=)
JP (2) JP7227901B2 (https=)
KR (1) KR102556363B1 (https=)
CN (1) CN110088624B (https=)
AU (1) AU2017345560C1 (https=)
BR (1) BR112019007831A2 (https=)
CA (1) CA3040507A1 (https=)
DK (1) DK3529616T5 (https=)
EA (1) EA039761B1 (https=)
ES (1) ES2963830T3 (https=)
FI (1) FI3529616T3 (https=)
HU (1) HUE064332T2 (https=)
MX (2) MX2019004431A (https=)
PL (1) PL3529616T3 (https=)
PT (1) PT3529616T (https=)
SA (1) SA519401607B1 (https=)
SG (1) SG11201903523YA (https=)
UA (1) UA129533C2 (https=)
WO (1) WO2018075783A2 (https=)
ZA (1) ZA201902654B (https=)

Families Citing this family (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2825370A1 (en) 2010-12-22 2012-06-28 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
WO2016077052A2 (en) 2014-10-22 2016-05-19 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
WO2017015545A1 (en) 2015-07-22 2017-01-26 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
WO2017019895A1 (en) 2015-07-30 2017-02-02 President And Fellows Of Harvard College Evolution of talens
IL310721B2 (en) 2015-10-23 2025-11-01 Harvard College Nucleobase editors and uses thereof
SG11201809903WA (en) * 2016-05-16 2018-12-28 Harvard College Method for purification and activation of botulinum neurotoxin
WO2018075783A2 (en) 2016-10-20 2018-04-26 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
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
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
EP4700040A1 (en) * 2023-04-21 2026-02-25 Shenzhen Loggene Life Science & Technology Co., Ltd. Peptide and luminescent complex containing same
GB202404021D0 (en) 2024-03-20 2024-05-01 Ipsen Biopharm Ltd Cell-based neurotoxin assay
KR20250158928A (ko) * 2024-04-30 2025-11-07 주식회사 바이오링크스 앵커 단백질을 이용한 미광성 세포기반의 보툴리눔 독소의 검출 및 활성 평가방법
KR20260002179A (ko) * 2024-06-28 2026-01-06 ㈜바이오솔빅스 Snap-25를 발현하는 운동신경세포 제조 방법 및 제조된 운동신경세포를 이용한 보툴리눔 신경독소 역가 측정 방법
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)

* 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
JP3523879B2 (ja) 1994-05-31 2004-04-26 アレルガン インコーポレイテッド 輸送タンパク質用クロストリジウム属細菌毒素の修飾
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
JP2002507410A (ja) 1998-03-27 2002-03-12 プロルーム・リミテッド ルシフェラーゼ、蛍光タンパク質、ルシフェラーゼおよび蛍光タンパク質をコードする核酸および、診断、高処理スクリーニングおよび新規アイテムにおけるその使用
US6890745B1 (en) 2000-07-19 2005-05-10 Chemicon International, Inc. Protease specific cleavable luciferases and methods of use thereof
CA2541765A1 (en) * 2003-10-10 2004-10-01 Promega Corporation Luciferase biosensor
US8753831B2 (en) 2007-06-05 2014-06-17 City Of Hope Methods for detection of botulinum neurotoxin
LT3409764T (lt) 2009-05-01 2020-06-10 Promega Corporation Sintetinės oplophorus liuciferazės su sustiprinta šviesos išeiga
US20140287433A1 (en) * 2011-07-19 2014-09-25 ETH Zürich Means and methods for determining clostridial neurotoxins
CA2866595A1 (en) 2012-03-07 2013-09-12 Merz Pharma Gmbh & Co. Kgaa Means and methods for determining neurotoxin activity based on a modified luciferase
CA2886939C (en) * 2012-10-16 2020-11-03 Merz Pharma Gmbh & Co. Kgaa Cellular test systems for the determination of the biological activities of neurotoxin polypeptides
KR20150087321A (ko) * 2012-11-21 2015-07-29 메르츠 파마 게엠베하 운트 코. 카가아 보툴리눔 신경독소 생물학적 활성의 결정을 위한 수단 및 방법
SG11201507306VA (en) 2013-03-15 2015-10-29 Promega Corp Activation of bioluminescence by structural complementation
AU2014301116B2 (en) * 2013-06-28 2019-11-14 Merz Pharma Gmbh & Co. Kgaa Means and methods for the determination of the biological activity of Neurotoxin polypeptides in cells
PT3274364T (pt) * 2015-03-26 2021-11-05 Harvard College Neurotoxina botulínica manipulada
WO2018075783A2 (en) 2016-10-20 2018-04-26 President And Fellows Of Harvard College In vitro and cell based assays for measuring the activity of botulinum neurotoxins
EP3893916A4 (en) * 2018-12-11 2023-03-29 Q32 Bio Inc. FUSION PROTEIN CONSTRUCTS FOR A COMPLEMENT-ASSOCIATED DISEASE

Also Published As

Publication number Publication date
PL3529616T3 (pl) 2024-03-18
ZA201902654B (en) 2024-08-28
JP7227901B2 (ja) 2023-02-22
US20220356508A1 (en) 2022-11-10
AU2017345560A1 (en) 2019-05-02
EP3529616A2 (en) 2019-08-28
EP3529616B1 (en) 2023-09-06
AU2017345560B2 (en) 2024-12-12
SG11201903523YA (en) 2019-05-30
KR102556363B1 (ko) 2023-07-18
MX2019004431A (es) 2019-08-14
HUE064332T2 (hu) 2024-03-28
DK3529616T3 (da) 2023-12-11
SA519401607B1 (ar) 2022-08-04
KR20190068582A (ko) 2019-06-18
EA039761B1 (ru) 2022-03-10
FI3529616T3 (fi) 2023-12-04
DK3529616T5 (da) 2024-09-02
MX2023004733A (es) 2023-05-10
WO2018075783A2 (en) 2018-04-26
ES2963830T3 (es) 2024-04-02
EA201990929A1 (ru) 2019-10-31
US11306347B2 (en) 2022-04-19
CN110088624B (zh) 2024-01-16
CN110088624A (zh) 2019-08-02
AU2017345560C1 (en) 2025-04-17
US11788069B2 (en) 2023-10-17
JP2019532651A (ja) 2019-11-14
JP2023011700A (ja) 2023-01-24
BR112019007831A2 (pt) 2019-10-01
UA129533C2 (uk) 2025-05-28
US20190276873A1 (en) 2019-09-12
PT3529616T (pt) 2023-12-06
WO2018075783A3 (en) 2018-06-21

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