DK2171098T3 - Fremgangsmåder til ekstraktion og rensning af komponenter i biologiske prøver - Google Patents

Fremgangsmåder til ekstraktion og rensning af komponenter i biologiske prøver Download PDF

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Publication number
DK2171098T3
DK2171098T3 DK08781182.4T DK08781182T DK2171098T3 DK 2171098 T3 DK2171098 T3 DK 2171098T3 DK 08781182 T DK08781182 T DK 08781182T DK 2171098 T3 DK2171098 T3 DK 2171098T3
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nucleic acid
elution
buffer
sample
neutralization
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DK08781182.4T
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English (en)
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Michael Lizzi
Matthew P Collis
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Becton Dickinson Co
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    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G01N33/54333—Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction
    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Processing Of Solid Wastes (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Claims (8)

1. Fremgangsmåde til ekstraktion af nucleinsyrekomponenter i en biologisk prøve, hvilken fremgangsmåde omfatter: (i) at binde reversibelt mindst en nucleinsyrekomponent i den biologiske prøve til mindst en paramagnetisk partikel i en beholder, hvilken paramagnetiske partikel er en jernoxidpartikel i et surt miljø; (ii) at adskille den mindst ene nucleinsyrekomponent, der er bundet til den mindst ene paramagnetiske partikel, fra ubundne komponenter i den biologiske prøve; (iii) at vaske den mindst ene nucleinsyrekomponent, der er bundet til den mindst ene paramagnetiske partikel; (iv) at adskille den mindst ene nucleinsyrekomponent, der er bundet til den mindst ene paramagnetiske partikel, fra vasken; (v) at fjerne den mindst ene nucleinsyrekomponent fra den mindst ene paramagnetiske partikel ved en eluerings- og neutraliseringsproces i to trin, hvilken proces omfatter: - at eluere den mindst ene nucleinsyrekomponent, der bundet til den mindst ene paramagnetiske partikel, med en opløsning af en elueringsbuffer med basisk pH, hvilken opløsning består af kaliumhydroxid eller natriumhydroxid, hvorved der opnås en elueret prøve; og - derefter at neutralisere den eluerede prøve ved efterfølgende at tilsætte en neutraliserende buffer, der er valgt blandt bicin, tris, 2-(cyclohexylamino)ethan-sul-fonsyre (CHES), N,N-bis(2-hydroxyethyl)-2-aminoethansulfonsyre (BES) , 4-morpholinpropansulfonsyre (MOPS) eller phosphat, hvorved der opnås en optimeret buffer, og (vi) derpå at adskille og fjerne de paramagnetiske partikler efter neutralisering af prøven, medens den pH-optimerede opløsning der indeholder den ubundne nuc-leinsyre overføres til yderligere bearbejdning, hvor fjernelsestrinnet er adskilt fra neutraliseringsstrinnet.
2. Fremgangsmåden ifølge krav 1, hvor den biologiske prøve er klinisk, retsmedicinsk eller miljømæssig prøve.
3. Fremgangsmåden ifølge krav 2, hvor den biologiske prøve er miljømæssig prøve, der omfatter jord, vand, luft, suspensionsudløb eller pulver.
4. Fremgangsmåden ifølge krav 1, hvor den biologiske prøve forbehandles til at lyse celler.
5. Fremgangsmåden ifølge krav 1, hvor elueringen omfatter at hæve pH med pH-elueringsbufferen.
6. Fremgangsmåden ifølge krav 1, hvor elueringsbufferen har en pH på 8 til 14.
7. Fremgangsmåden ifølge krav 1, hvor neutraliseringsbufferen sænker pH.
8. Fremgangsmåden ifølge krav 7, hvor pH er 6 til 9, fortrinsvis hvor pH er 8 til 8,5, mere foretrukket, hvor pH er 8,4 efter tilsætningen af neutraliserings-bufferen.
DK08781182.4T 2007-06-29 2008-06-30 Fremgangsmåder til ekstraktion og rensning af komponenter i biologiske prøver DK2171098T3 (da)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US92951207P 2007-06-29 2007-06-29
US92954407P 2007-07-02 2007-07-02
PCT/US2008/068807 WO2009006417A2 (en) 2007-06-29 2008-06-30 Methods for extraction and purification of components of biological samples

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DK2171098T3 true DK2171098T3 (da) 2018-05-22

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DK08781182.4T DK2171098T3 (da) 2007-06-29 2008-06-30 Fremgangsmåder til ekstraktion og rensning af komponenter i biologiske prøver

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US (2) US20090061497A1 (da)
EP (1) EP2171098B1 (da)
JP (1) JP5232858B2 (da)
AU (1) AU2008273030B2 (da)
CA (1) CA2692186C (da)
DK (1) DK2171098T3 (da)
ES (1) ES2665280T3 (da)
WO (1) WO2009006417A2 (da)

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* Cited by examiner, † Cited by third party
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WO2022059762A1 (ja) * 2020-09-18 2022-03-24 国立大学法人東海国立大学機構 生体分子の抽出方法

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WO2009006417A3 (en) 2009-02-26
US20170191054A1 (en) 2017-07-06
CA2692186C (en) 2019-03-12
US20090061497A1 (en) 2009-03-05
EP2171098A4 (en) 2010-11-03
JP5232858B2 (ja) 2013-07-10
EP2171098A2 (en) 2010-04-07
AU2008273030B2 (en) 2013-01-17
JP2010532483A (ja) 2010-10-07
CA2692186A1 (en) 2009-01-08
AU2008273030A1 (en) 2009-01-08
EP2171098B1 (en) 2018-03-28
WO2009006417A2 (en) 2009-01-08
ES2665280T3 (es) 2018-04-25

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