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1. WO2020066336 - DETECTION METHOD AND ASSESSMENT METHOD

Publication Number WO/2020/066336
Publication Date 02.04.2020
International Application No. PCT/JP2019/031419
International Filing Date 08.08.2019
IPC
G01N 33/53 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
G01N 21/64 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
G01N 33/536 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
536with immune complex formed in liquid phase
G01N 33/541 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
536with immune complex formed in liquid phase
537with separation of immune complex from unbound antigen or antibody
539involving precipitating reagent
541Double or second antibody
C12Q 1/02 2006.01
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
02involving viable microorganisms
CPC
C12Q 1/02
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
02involving viable microorganisms
G01N 21/64
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
G01N 33/53
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
G01N 33/536
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
536with immune complex formed in liquid phase
G01N 33/541
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
536with immune complex formed in liquid phase
537with separation of immune complex from unbound antigen or antibody
539involving precipitating reagent ; , e.g. ammonium sulfate
541Double or second antibody ; , i.e. precipitating antibody
Applicants
  • コニカミノルタ株式会社 KONICA MINOLTA, INC. [JP]/[JP]
Inventors
  • 相宮 拓司 AIMIYA Takuji
  • 古澤 直子 FURUSAWA Naoko
  • 中野 寧 NAKANO Yasushi
  • 権田 幸祐 GONDA Kohsuke
  • 北村 成史 KITAMURA Narufumi
Agents
  • 特許業務法人SSINPAT SSINPAT PATENT FIRM
Priority Data
2018-18041826.09.2018JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) DETECTION METHOD AND ASSESSMENT METHOD
(FR) PROCÉDÉ DE DÉTECTION ET D'ÉVALUATION
(JA) 検出方法および評価方法
Abstract
(EN)
The present invention provides a method for detecting a protein complex by causing blinking of a fluorescent particle, which does not exhibit blinking, due to energy transfer from a fluorescent particle which exhibits excited blinking. The detection method for a protein complex of a protein (A) and a protein (B) according to the present invention, comprises a step (a) for labeling the protein (A) with an antibody to which a fluorescent particle which exhibits blinking is linked, a step (b) for labeling the protein (B) with an antibody to which a fluorescent particle which does not exhibit blinking is linked, and a step (c) for, after steps (a) and (b), irradiating, with light having an excitation wavelength, the fluorescent particle which exhibits blinking, at the same time, detecting bright points of fluorescence of the emission wavelength of the fluorescent particles which do not exhibit blinking, and extracting a blinking bright point from the bright points of fluorescence of the emission wavelength of the fluorescent particles which do not exhibit blinking.
(FR)
La présente invention concerne un procédé de détection d'un complexe protéique en provoquant le clignotement d'une particule fluorescente, qui ne présente pas de clignotement, grâce au transfert d'énergie d'une particule fluorescente qui présente un clignotement excité. Selon la présente invention, le procédé de détection d'un complexe protéique d'une protéine (A) et d'une protéine (B) comprend une étape (a) où la protéine (A) est marquée avec un anticorps auquel une particule fluorescente qui présente un clignotement est liée, une étape (b) où la protéine (B) est marquée avec un anticorps auquel une particule fluorescente qui ne présente pas de clignotement est liée, et après les étapes (a) et (b), une étape (c) pour irradier, avec la lumière ayant une longueur d'onde d'excitation, la particule fluorescente qui présente un clignotement et détecter parallèlement des points lumineux de fluorescence de la longueur d'onde d'émission à partir des particules fluorescentes qui ne présentent pas de clignotement, et extraire un point lumineux clignotant des points lumineux de fluorescence de la longueur d'onde d'émission à partir des particules fluorescentes qui ne présentent pas de clignotement.
(JA)
本発明は、励起されたブリンキングを生じる蛍光発光性粒子からのエネルギー移動によりブリンキングを生じない蛍光発光性粒子を明滅させることでタンパク質複合体を検出する方法を提供する。本発明に係るタンパク質(A)とタンパク質(B)とからなるタンパク質複合体の検出方法は、タンパク質(A)を、ブリンキングを生じる蛍光発光性粒子を結合した抗体を用いて標識する工程(a)、およびタンパク質(B)を、ブリンキングを生じない蛍光発光性粒子を結合した抗体で標識する工程(b)を行った後、ブリンキングを生じる蛍光発光性粒子の励起波長の光を照射すると同時に、ブリンキングを生じない蛍光発光性粒子の発光波長の蛍光輝点を検出し、前記ブリンキングを生じない蛍光発光性粒子の発光波長の蛍光輝点から、明滅する輝点を抽出する工程(c)を含む。
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