Fluorescence polarization assay for determining histidine...

Details Fluorescence polarization assay for determining histidine... Fluorescence polarization assay for determining histidine...

G - Physics

01

N

G01N 33/58 (2006.01) G01N 33/53 (2006.01) G01N 33/533 (2006.01) G01N 33/574 (2006.01)

Patent

CA 2582135

A fluorescence polarization assay for determining the HDC modulating activity of a candidate compound comprising the steps of: a) providing a reaction mixture comprising a HDC, histidine, a fluorescently labeled histamine probe, a candidate compound and an anti histidine antibody having selectivity for histamine at least 10 fold greater than histidine; b) incubating the reaction mixture; c) determining whether inhibition of HDC has occured in the presence of the test compound, wherein an increase in fluorescence signal is an indication that the test compound inhibits the activity of HDC.

L~invention concerne une analyse par polarisation de fluorescence visant à déterminer l~activité de modulation de la HDC d~un composé candidat, ladite analyse comprenant les étapes consistant à : a) préparer un mélange réactionnel comprenant une HDC, de l~histidine, une sonde à histamine marquée de manière fluorescente, un composé candidat et un anticorps anti-histidine présentant une sélectivité pour l~histamine au moins dix fois supérieure à l~histidine ; b) incuber le mélange réactionnel ; c) déterminer si l~inhibition de la HDC est survenue en présence du composé test. Une augmentation du signal fluorescent est une indication de l~inhibition de l~activité de la HDC par le composé test.

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