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1. WO2020138265 - METHOD, APPARATUS AND PROGRAM FOR ASSESSING AMYLOID FORMATION

Publication Number WO/2020/138265
Publication Date 02.07.2020
International Application No. PCT/JP2019/051077
International Filing Date 26.12.2019
IPC
G01N 21/64 2006.1
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 33/15 2006.1
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
15Medicinal preparations
G01N 33/50 2006.1
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
CPC
C07K 14/47
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
435from animals; from humans
46from vertebrates
47from mammals
C07K 16/18
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
16Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
18against material from animals or humans
C12M 1/34
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
1Apparatus for enzymology or microbiology
34Measuring or testing with condition measuring or sensing means, e.g. colony counters
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 sub-millimetre waves, infrared, visible or ultraviolet 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 33/15
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
15Medicinal preparations ; ; Physical properties thereof, e.g. dissolubility
G01N 33/50
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
Applicants
  • 国立大学法人室蘭工業大学 NATIONAL UNIVERSITY CORPORATION MURORAN INSTITUTE OF TECHNOLOGY [JP]/[JP]
Inventors
  • 徳楽 清孝 TOKURAKU Kiyotaka
  • 上井 幸司 UWAI Koji
Agents
  • 特許業務法人IPアシスト特許事務所 IP-ASSIST PATENT OFFICE
Priority Data
2018-24228026.12.2018JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) METHOD, APPARATUS AND PROGRAM FOR ASSESSING AMYLOID FORMATION
(FR) PROCÉDÉ, APPAREIL ET PROGRAMME POUR ÉVALUER LA FORMATION D'AMYLOÏDE
(JA) アミロイド形成を評価する方法、装置及びプログラム
Abstract
(EN)
The present invention pertains to: a method for determining amyloid formation inhibitory activities of test substances, the method comprising: a step for reacting, in the presence or absence of a test substance, an amyloidogenic protein with a fluorescent probe capable of binding to amyloid formed by polymerizing the amyloidogenic protein; a step for capturing the fluorescence of agglutination reaction products under an exposure condition in which Sum Intensity calculated from the brightness value of each pixel included in a region of interest in a fluorescence image to be captured is 15-85%; a step for calculating the standard deviation from the brightness value of each pixel included in the region of interest in the captured fluorescence image; and a step for comparing the calculated standard deviation in brightness values in the presence of the test substance and the standard deviation in brightness values in the absence of the test substance, and determining that the test substance has an amyloid formation inhibitory activity when the standard deviation in brightness values in the presence of the test substance is smaller than the standard deviation in brightness values in the absence of the test substance; and an apparatus and a program which can be used in implementing the method.
(FR)
La présente invention concerne : un procédé consistant à déterminer des activités inhibitrices de formation d'amyloïde de substances de test, le procédé comprenant : une étape consistant à faire réagir, en présence ou en l'absence d'une substance de test, une protéine amyloïdogène avec une sonde fluorescente capable de se lier à l'amyloïde formée par polymérisation de la protéine amyloïdogène ; une étape consistant à capturer la fluorescence des produits de réaction d'agglutination dans une condition d'exposition dans laquelle une intensité de somme calculée à partir de la valeur de luminosité de chaque pixel inclus dans une région d'intérêt dans une image de fluorescence devant être capturée est de 15 à 85 % ; une étape consistant à calculer l'écart type à partir de la valeur de luminosité de chaque pixel inclus dans la région d'intérêt dans l'image de fluorescence capturée ; et une étape consistant à comparer l'écart-type calculé dans des valeurs de luminosité en présence de la substance de test et de l'écart-type dans des valeurs de luminosité en l'absence de la substance de test, et à déterminer que la substance de test a une activité inhibitrice de formation d'amyloïde lorsque l'écart-type dans les valeurs de luminosité en présence de la substance de test est inférieur à l'écart-type dans des valeurs de luminosité en l'absence de la substance de test ; et un appareil et un programme qui peuvent être utilisés dans la mise en œuvre du procédé.
(JA)
本発明は、被験物質の存在下又は非存在下でアミロイド形成性タンパク質と前記アミロイド形成性タンパク質の重合により形成されるアミロイドへの結合能を有する蛍光プローブとを反応させるステップと;凝集反応生成物の蛍光を撮像される蛍光画像内の関心領域に含まれる各画素の輝度値から算出されるSum Intensityが15~85%となる露出条件で撮像するステップと;撮像した蛍光画像内の関心領域に含まれる各画素の輝度値から標準偏差を算出するステップと;算出した被験物質存在下での輝度値標準偏差の値と被験物質非存在下での輝度値標準偏差の値とを比較し、被験物質存在下での輝度値標準偏差の値が被験物質非存在下での輝度値標準偏差の値より小さい場合に被験物質はアミロイド形成阻害活性を有すると判定するステップとを含む、被験物質のアミロイド形成阻害活性を判定する方法、並びに前記方法の実施において使用可能な装置及びプログラムに関する。
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