WO2010111686A3 - Labeled enzyme compositions, methods & systems - Google Patents

Labeled enzyme compositions, methods & systems Download PDF

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Publication number
WO2010111686A3
WO2010111686A3 PCT/US2010/028967 US2010028967W WO2010111686A3 WO 2010111686 A3 WO2010111686 A3 WO 2010111686A3 US 2010028967 W US2010028967 W US 2010028967W WO 2010111686 A3 WO2010111686 A3 WO 2010111686A3
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WO
WIPO (PCT)
Prior art keywords
conjugates
methods
systems
polymerase
enzyme compositions
Prior art date
Application number
PCT/US2010/028967
Other languages
French (fr)
Other versions
WO2010111686A2 (en
Inventor
Theo Nikiforov
Original Assignee
Life Technologies Corp
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Publication date
Application filed by Life Technologies Corp filed Critical Life Technologies Corp
Publication of WO2010111686A2 publication Critical patent/WO2010111686A2/en
Publication of WO2010111686A3 publication Critical patent/WO2010111686A3/en

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    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
    • 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
    • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/07—Nucleotidyltransferases (2.7.7)
    • C12Y207/07007—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • 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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/07—Nucleotidyltransferases (2.7.7)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6432—Quenching

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • General Physics & Mathematics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Disclosed herein are conjugates comprising a biomolecule linked to a label that have biological activity and are useful in a wide variety of biological applications. For example, provided herein are labeled polymerase conjugates including a polymerase linked to one or more labels, wherein the conjugate has polymerase activity. Such conjugates can exhibit enhanced biological activity and/or superior detectability as compared to conventional labeled polymerases. Also disclosed herein are improved methods for preparing such conjugates, and methods and systems for using such conjugates in biological applications such as nucleotide incorporation, primer extension and single molecule sequencing.
PCT/US2010/028967 2009-03-27 2010-03-26 Labeled enzyme compositions, methods & systems WO2010111686A2 (en)

Applications Claiming Priority (22)

Application Number Priority Date Filing Date Title
US16432409P 2009-03-27 2009-03-27
US61/164,324 2009-03-27
US18477009P 2009-06-05 2009-06-05
US61/184,770 2009-06-05
US24277109P 2009-09-15 2009-09-15
US61/242,771 2009-09-15
US24545709P 2009-09-24 2009-09-24
US61/245,457 2009-09-24
US26397409P 2009-11-24 2009-11-24
US61/263,974 2009-11-24
US28938809P 2009-12-22 2009-12-22
US61/289,388 2009-12-22
US29361610P 2010-01-08 2010-01-08
US29361810P 2010-01-08 2010-01-08
US61/293,616 2010-01-08
US61/293,618 2010-01-08
US29991910P 2010-01-29 2010-01-29
US29991710P 2010-01-29 2010-01-29
US61/299,917 2010-01-29
US61/299,919 2010-01-29
US30735610P 2010-02-23 2010-02-23
US61/307,356 2010-02-23

Publications (2)

Publication Number Publication Date
WO2010111686A2 WO2010111686A2 (en) 2010-09-30
WO2010111686A3 true WO2010111686A3 (en) 2011-05-19

Family

ID=42781937

Family Applications (4)

Application Number Title Priority Date Filing Date
PCT/US2010/028967 WO2010111686A2 (en) 2009-03-27 2010-03-26 Labeled enzyme compositions, methods & systems
PCT/US2010/028972 WO2010111690A2 (en) 2009-03-27 2010-03-26 Polymerase compositions and methods
PCT/US2010/028974 WO2010111691A2 (en) 2009-03-27 2010-03-26 Conjugates of biomolecules to nanoparticles
PCT/US2010/028952 WO2010111674A2 (en) 2009-03-27 2010-03-26 Methods and apparatus for single molecule sequencing using energy transfer detection

Family Applications After (3)

Application Number Title Priority Date Filing Date
PCT/US2010/028972 WO2010111690A2 (en) 2009-03-27 2010-03-26 Polymerase compositions and methods
PCT/US2010/028974 WO2010111691A2 (en) 2009-03-27 2010-03-26 Conjugates of biomolecules to nanoparticles
PCT/US2010/028952 WO2010111674A2 (en) 2009-03-27 2010-03-26 Methods and apparatus for single molecule sequencing using energy transfer detection

Country Status (2)

Country Link
US (20) US8603792B2 (en)
WO (4) WO2010111686A2 (en)

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US8603792B2 (en) 2009-03-27 2013-12-10 Life Technologies Corporation Conjugates of biomolecules to nanoparticles

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WO2010111690A3 (en) 2011-03-31
WO2010111674A3 (en) 2011-05-12
US20230055565A1 (en) 2023-02-23
US20110014612A1 (en) 2011-01-20
US20150118679A1 (en) 2015-04-30
US20160186243A1 (en) 2016-06-30
WO2010111691A9 (en) 2011-01-06
US10093974B2 (en) 2018-10-09
US11008612B2 (en) 2021-05-18
WO2010111686A2 (en) 2010-09-30
US20190062830A1 (en) 2019-02-28
US20190062828A1 (en) 2019-02-28
US20120322057A1 (en) 2012-12-20
US9932573B2 (en) 2018-04-03
US20110003343A1 (en) 2011-01-06
US8999674B2 (en) 2015-04-07
US11015220B2 (en) 2021-05-25
US20170191051A1 (en) 2017-07-06
US20130040363A1 (en) 2013-02-14
US20100261185A1 (en) 2010-10-14
US11542549B2 (en) 2023-01-03
WO2010111674A2 (en) 2010-09-30
US20160265045A1 (en) 2016-09-15
US9365838B2 (en) 2016-06-14
US20170283868A1 (en) 2017-10-05
US20180258415A1 (en) 2018-09-13
US20100255487A1 (en) 2010-10-07
US20120329042A1 (en) 2012-12-27
US10093972B2 (en) 2018-10-09
US20150247136A1 (en) 2015-09-03
US9365839B2 (en) 2016-06-14
US9567629B2 (en) 2017-02-14

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