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MX2016005270A - Ensayo enzimatico con fluoroforos duplicados. - Google Patents

Ensayo enzimatico con fluoroforos duplicados.

Info

Publication number
MX2016005270A
MX2016005270A MX2016005270A MX2016005270A MX2016005270A MX 2016005270 A MX2016005270 A MX 2016005270A MX 2016005270 A MX2016005270 A MX 2016005270A MX 2016005270 A MX2016005270 A MX 2016005270A MX 2016005270 A MX2016005270 A MX 2016005270A
Authority
MX
Mexico
Prior art keywords
region
fluorophores
duplicate
enzyme assay
cleavage site
Prior art date
Application number
MX2016005270A
Other languages
English (en)
Inventor
C Tucker Ward
Original Assignee
Biomadison Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Biomadison Inc filed Critical Biomadison Inc
Publication of MX2016005270A publication Critical patent/MX2016005270A/es

Links

Classifications

    • 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
    • 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
    • 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
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/50—Fusion polypeptide containing protease site
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/95—Fusion polypeptide containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence)
    • 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
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
    • G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
    • G01N2333/952—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from bacteria

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Biophysics (AREA)
  • Toxicology (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Cell Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Se describen composiciones y métodos que proporcionan un ensayo a base de células rápido, sensible y exacto para la actividad enzimática, en particular para las actividades enzimáticas asociadas con las toxinas botulínicas. Se proporciona una célula que expresa una construcción que incluye una región de anclaje, un sitio de escisión y una región de reporte que tiene dos o más péptidos reporteros idénticos. La actividad enzimática en el sitio de escisión libera la región reportera en el citosol, donde se producen varios eventos de degradación. El cambio observado en la señal es proporcional a la actividad enzimática.
MX2016005270A 2013-10-25 2014-10-24 Ensayo enzimatico con fluoroforos duplicados. MX2016005270A (es)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US201361895533P 2013-10-25 2013-10-25
US201361897352P 2013-10-30 2013-10-30
US201462014586P 2014-06-19 2014-06-19
US201462058532P 2014-10-01 2014-10-01
PCT/US2014/062260 WO2015061738A1 (en) 2013-10-25 2014-10-24 Enzyme assay with duplicate fluorophores

Publications (1)

Publication Number Publication Date
MX2016005270A true MX2016005270A (es) 2017-02-02

Family

ID=52001049

Family Applications (2)

Application Number Title Priority Date Filing Date
MX2019013952A MX377925B (es) 2013-10-25 2014-10-24 Ensayo enzimatico con fluoroforos duplicados.
MX2016005270A MX2016005270A (es) 2013-10-25 2014-10-24 Ensayo enzimatico con fluoroforos duplicados.

Family Applications Before (1)

Application Number Title Priority Date Filing Date
MX2019013952A MX377925B (es) 2013-10-25 2014-10-24 Ensayo enzimatico con fluoroforos duplicados.

Country Status (13)

Country Link
US (3) US20150118701A1 (es)
EP (2) EP3404112B1 (es)
JP (2) JP6636919B2 (es)
KR (1) KR20160103977A (es)
CN (2) CN113073129A (es)
AU (1) AU2014339884B2 (es)
CA (1) CA2928609C (es)
ES (2) ES2829612T3 (es)
LT (2) LT3404112T (es)
MX (2) MX377925B (es)
RU (1) RU2683207C2 (es)
TW (1) TWI647237B (es)
WO (1) WO2015061738A1 (es)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3030905B9 (en) * 2013-08-09 2024-04-03 Biomadison, Inc. Botulinum toxin assay with improved sensitivity
EP3404112B1 (en) * 2013-10-25 2020-08-26 BioMadison, Inc. Enzyme assay with duplicate fluorophores
US10441640B2 (en) * 2014-08-12 2019-10-15 Biomadison, Inc. Botulinum neurotoxins with modified light chain specificity and methods for producing same
US12019066B2 (en) * 2016-05-16 2024-06-25 Biomadison, Inc. Assay with synaptobrevin based moiety
EP3458600A4 (en) 2016-05-16 2020-01-15 BioMadison, Inc. Improved assay with synaptobrevin based moiety
US11396499B2 (en) 2018-12-12 2022-07-26 University Of Washington Lysosomal acid lipase assay
TW202248635A (zh) * 2021-04-06 2022-12-16 美商伯克利之光生命科技公司 微流體分析與來自非哺乳動物細胞之生物生產的方法及用於其之套組

Family Cites Families (19)

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AU759180B2 (en) * 1998-10-30 2003-04-10 Cellomics, Inc. A system for cell-based screening
DE10038382A1 (de) * 2000-08-07 2002-02-21 Direvo Biotech Ag Zweifarbiges fluorimetrisches Proteaseassay
EP1349867A4 (en) 2000-12-15 2004-10-27 Stratagene California DIMER FLUORESCENT POLYPEPTIDES
US7183066B2 (en) 2002-09-27 2007-02-27 Allergan, Inc. Cell-based fluorescence resonance energy transfer (FRET) assays for clostridial toxins
JP4214206B2 (ja) * 2002-12-10 2009-01-28 独立行政法人理化学研究所 Ｆｒｅｔを利用した蛍光指示薬
US7667024B2 (en) * 2003-11-19 2010-02-23 Allelogic Biosciences Corp. Oligonucleotides labeled with a plurality of fluorophores
EP2502930B1 (en) * 2003-12-19 2016-06-29 Wisconsin Alumni Research Foundation Method and compositions for detecting botulinum neurotoxin
WO2006107864A1 (en) * 2005-04-04 2006-10-12 Blueshift Biotechnologies, Inc. Screening using polarization anisotropy in fret emissions
ES2351942T3 (es) * 2005-04-05 2011-02-14 Allergan Inc Analisis de la actividad de una toxina clostridial.
CA2622422A1 (en) * 2005-09-15 2007-03-29 Mark Mercola Methods for screening for compounds that modulate insulin promoter activity
KR100883158B1 (ko) * 2005-10-26 2009-02-10 주식회사 뉴젝스 살아있는 세포에서 단백질분해효소의 활성 측정 방법
EP1997885A1 (en) 2007-05-29 2008-12-03 AXXAM S.p.A. Fluorescent proteins and methods of use thereof
US8753831B2 (en) * 2007-06-05 2014-06-17 City Of Hope Methods for detection of botulinum neurotoxin
JP5826635B2 (ja) * 2009-03-13 2015-12-02 アラーガン、インコーポレイテッドＡｌｌｅｒｇａｎ，Ｉｎｃｏｒｐｏｒａｔｅｄ 免疫系調節エンドペプチターゼ活性アッセイ
US10246492B2 (en) 2009-10-16 2019-04-02 Biomadison, Inc. Botulinum assay with synaptobrevin substrate moiety
WO2011047265A1 (en) * 2009-10-16 2011-04-21 Biosentinel, Inc. Resonance energy transfer assay with synaptobrevin substrate moiety
CN103842819B (zh) 2011-06-01 2017-09-12 拜奥麦迪逊公司 非‑fret肉毒梭菌测定法
EP3030905B9 (en) 2013-08-09 2024-04-03 Biomadison, Inc. Botulinum toxin assay with improved sensitivity
EP3404112B1 (en) * 2013-10-25 2020-08-26 BioMadison, Inc. Enzyme assay with duplicate fluorophores

Also Published As

Publication number Publication date
US20210032675A1 (en) 2021-02-04
ES2686141T3 (es) 2018-10-16
MX2019013952A (es) 2020-01-30
JP2016533733A (ja) 2016-11-04
WO2015061738A1 (en) 2015-04-30
WO2015061738A4 (en) 2015-07-02
KR20160103977A (ko) 2016-09-02
US20150118701A1 (en) 2015-04-30
RU2016118286A (ru) 2017-11-30
RU2683207C2 (ru) 2019-03-26
LT3060677T (lt) 2018-08-27
US11685945B2 (en) 2023-06-27
CA2928609C (en) 2022-03-15
TW201531481A (zh) 2015-08-16
CA2928609A1 (en) 2015-04-30
RU2016118286A3 (es) 2018-05-21
EP3060677B1 (en) 2018-07-04
ES2829612T3 (es) 2021-06-01
JP2020062032A (ja) 2020-04-23
CN106414763A (zh) 2017-02-15
US20240117408A1 (en) 2024-04-11
LT3404112T (lt) 2020-12-28
EP3060677A1 (en) 2016-08-31
AU2014339884A1 (en) 2016-05-12
EP3404112A1 (en) 2018-11-21
CN113073129A (zh) 2021-07-06
CN106414763B (zh) 2021-04-09
JP6636919B2 (ja) 2020-01-29
EP3404112B1 (en) 2020-08-26
MX377925B (es) 2025-03-10
US12209269B2 (en) 2025-01-28
TWI647237B (zh) 2019-01-11
AU2014339884B2 (en) 2020-03-05

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