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1. WO2009131948 - HIGH-THROUGHPUT CELL-BASED CFTR ASSAY

Publication Number WO/2009/131948
Publication Date 29.10.2009
International Application No. PCT/US2009/041147
International Filing Date 20.04.2009
IPC
C12Q 1/68 2006.1
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
CPC
G01N 33/5038
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
5005involving human or animal cells
5008for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
502for testing non-proliferative effects
5038involving detection of metabolites per se
G01N 33/5067
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
5005involving human or animal cells
5008for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
5044involving specific cell types
5067Liver cells
G01N 33/6872
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
68involving proteins, peptides or amino acids
6872Intracellular protein regulatory factors and their receptors, e.g. including ion channels
Applicants
  • INSTITUTE FOR ONEWORLD HEALTH [US]/[US] (AllExceptUS)
  • GARDENER, Matthew, James [GB]/[GB] (UsOnly)
  • SHEARER, Joanne, Lesley [GB]/[GB] (UsOnly)
  • CRONK, David, William [GB]/[GB] (UsOnly)
Inventors
  • GARDENER, Matthew, James
  • SHEARER, Joanne, Lesley
  • CRONK, David, William
Agents
  • KONSKI, Antoinette, F.
Priority Data
61/046,74621.04.2008US
Publication Language English (en)
Filing Language English (EN)
Designated States
Title
(EN) HIGH-THROUGHPUT CELL-BASED CFTR ASSAY
(FR) DOSAGE À BASE CELLULAIRE ET À HAUT RENDEMENT DE LA CFTR
Abstract
(EN) This invention provides a high throughput screen for measuring the transport of an ion through a cystic fibrosis transmembrane conductance regulator (CFTR) of a cell that endogenously expresses CFTR. The method requires culturing the cell that endogenously expresses CFTR in the presence of an ion-sensitive compound and then contacting the cell with a CFTR activator. After a suitable amount of time after addition of the activator, the test compound is added to the cell culture medium. Thereafter, any change in the ion-sensitive compound is measured by any suitable method that can detect the change in the ion-sensitive compound, i.e. by measuring a change in the light emitted from a fluorescent compound, which can be recorded by an imaging plate reader.
(FR) La présente invention concerne un criblage à haut débit permettant la mesure du transport d'un ion par un régulateur de la conductance transmembranaire de la mucoviscidose (CFTR) d'une cellule exprimant, de façon endogène, ledit CFTR. Ce procédé implique la culture de la cellule exprimant de façon endogène le CFTR en présence d'un composé sensible aux ions, cela étant suivi de la mise en contact de la cellule avec un activateur du CFTR. A l'issue d'un laps de temps suffisant après l'addition de l'activateur, le composé à étudier est ajouté au milieu de culture de la cellule. Par la suite, toute modification affectant le composé sensible aux ions est mesurée par un quelconque procédé adapté capable de détecter ladite modification affectant le composé sensible aux ions, par exemple grâce à la mesure d'une modification affectant la lumière émise par un composé fluorescent, qui peut être enregistrée par un lecteur de plaques d'imagerie.
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