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1. WO2020067121 - ALKALINE PHOSPHATASE COMPOSITION, METHOD FOR PRODUCING DEPHOSPHORYLATED NUCLEIC ACID AND METHOD FOR PRODUCING LABELED NUCLEIC ACID

Publication Number WO/2020/067121
Publication Date 02.04.2020
International Application No. PCT/JP2019/037519
International Filing Date 25.09.2019
IPC
C12N 9/16 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14Hydrolases (3.)
16acting on ester bonds (3.1)
C12N 15/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
C12N 15/55 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
52Genes encoding for enzymes or proenzymes
55Hydrolases (3)
C12Q 1/6874 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6869Methods for sequencing
6874involving nucleic acid arrays, e.g. sequencing by hybridisation
CPC
C12N 15/10
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
C12N 9/16
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
16acting on ester bonds (3.1)
C12Q 1/6874
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6869Methods for sequencing
6874involving nucleic acid arrays, e.g. sequencing by hybridisation
Applicants
  • 東レ株式会社 TORAY INDUSTRIES, INC. [JP]/[JP]
Inventors
  • 伊藤 正照 ITO Masateru
  • 上田 洋二 UEDA Yoji
  • 瀧井 有樹 TAKII Yuki
  • 八木 麻衣 YAGI Mai
Agents
  • 永井 浩之 NAGAI Hiroshi
  • 中村 行孝 NAKAMURA Yukitaka
  • 朝倉 悟 ASAKURA Satoru
  • 浅野 真理 ASANO Makoto
  • 反町 洋 SORIMACHI Hiroshi
  • 鈴木 啓靖 SUZUKI Hiroyasu
Priority Data
2018-17953325.09.2018JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) ALKALINE PHOSPHATASE COMPOSITION, METHOD FOR PRODUCING DEPHOSPHORYLATED NUCLEIC ACID AND METHOD FOR PRODUCING LABELED NUCLEIC ACID
(FR) COMPOSITION DE PHOSPHATASE ALCALINE, PROCÉDÉ DE PRODUCTION D'ACIDE NUCLÉIQUE DÉPHOSPHORYLÉ ET PROCÉDÉ DE PRODUCTION D'ACIDE NUCLÉIQUE MARQUÉ
(JA) アルカリホスファターゼ組成物並びに脱リン酸化核酸及び標識化核酸の製造方法
Abstract
(EN)
The purpose of the present invention is to provide a high-quality composition containing alkaline phosphatase, a method for producing a dephosphorylated nucleic acid and a method for producing a labeled nucleic acid, each method comprising using the aforesaid composition. To achieve this purpose, provided is a composition that contains alkaline phosphatase and first to sixth peptide fragments, wherein the ratios of the contents of the first to sixth peptide fragments to the content of the alkaline phosphatase respectively satisfy the following requirements (1) to (6): (X1/Y)×100≤0.6000 (1); (X2/Y)×100≤0.1800 (2); (X3/Y)×100≤0.2000 (3); (X4/Y)×100≤0.8000 (4); (X5/Y)×100≤1.6000 (5); and (X6/Y)×100≤0.3500 (6) [wherein X1 to X6 respectively represent peak area values of the first to sixth peptide fragments which are calculated by the automatic integration method from an extraction ion chromatogram obtained by analyzing the composition by LC-MS/MS, and Y represents the peak area value of the alkaline phosphatase which is calculated by the automatic integration method from a chromatogram obtained by analyzing the composition by LC-UV].
(FR)
L'objectif de la présente invention est de fournir une composition de haute qualité contenant une phosphatase alcaline, un procédé de production d'un acide nucléique déphosphorylé et un procédé de production d'un acide nucléique marqué, chaque procédé comprenant l'utilisation de la composition susmentionnée. À cet effet, l'invention concerne une composition qui contient une phosphatase alcaline et les fragments peptidiques du premier au sixième, les rapports des teneurs des fragments peptidiques premier à sixième au contenu de la phosphatase alcaline satisfaisant respectivement les exigences (1) à (6) suivantes : (X1/Y)×100≤0.6000 (1) ; (X2/Y)×100≤0.1800 (2) ; (X3/Y)×100≤0.2000 (3) ; (X4/Y)×100≤0.8000 (4) ; (X5/Y)×100≤1.6000 (5) ; et (X6/Y)×100≤0.3500 (6) [dans laquelle X1 à X6 représentent respectivement des valeurs de zone de pic des fragments peptidiques premier à sixième qui sont calculées par le procédé d'intégration automatique à partir d'un chromatogramme d'ions d'extraction obtenu par analyse de la composition par couplage chromatographie liquide-spectrométrie de masse/spectrométrie de masse (ou LC-MS/MS) et Y représente la valeur de zone de pic de la phosphatase alcaline qui est calculée par le procédé d'intégration automatique à partir d’un chromatogramme obtenu par analyse de la composition par couplage chromatographie liquide-détecteur UV (ou LC-UV)].
(JA)
本発明は、アルカリホスファターゼを含有する高品質の組成物並びに該組成物を使用した脱リン酸化核酸の製造方法及び標識化核酸の製造方法を提供することを目的とし、かかる目的を達成するために、本発明は、アルカリホスファターゼと、第1~第6のペプチド断片とを含有する組成物であって、前記アルカリホスファターゼの含有量に対する前記第1~第6のペプチド断片の含有量の比率が、それぞれ、下記式(1)~(6): (X/Y)×100≦0.6000 ・・・(1) (X/Y)×100≦0.1800 ・・・(2) (X/Y)×100≦0.2000 ・・・(3) (X/Y)×100≦0.8000 ・・・(4) (X/Y)×100≦1.6000 ・・・(5) (X/Y)×100≦0.3500 ・・・(6) [式中、X~Xは、それぞれ、前記組成物のLC-MS/MS分析により得られた抽出イオンクロマトグラムから自動積分法により算出された、前記第1~第6のペプチド断片のピーク面積値を表し、Yは、前記組成物のLC-UV分析により得られたクロマトグラムから自動積分法により算出された、前記アルカリホスファターゼのピーク面積値を表す。] を満たす、前記組成物を提供する。
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