Multiplexed multitarget screening method

G - Physics – 01 – N

Patent

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G01N 15/14 (2006.01) C12M 1/34 (2006.01) C12M 3/02 (2006.01) G01N 33/53 (2006.01)

Patent

CA 2524782

The present disclosure provides systems for multiplexed multitarget screening of cell populations having one or more wild type or mutated ligand targets and measuring cell responses to ligands using high throughput screening techniques, including flow cytometry (FCM). The method includes the steps of: 1) developing cell populations to be screened; 2) staining cell populations using one or more fluorochromes to yield a distinct excitation/emission signature for each cell population; 3) combining labelled cell populations into a single mixed suspension; 4) analyzing populations to resolve them on the basis of their unique signature; and 5) resolving individual populations and deconvoluting data to extract meaningful information about populations.

L'invention concerne des systèmes de criblage mutiplexé multicible de populations de cellules possédant un ou plusieurs types sauvages ou des cibles de ligands mutants et permettant de mesurer les réponses des cellules à des ligands, au moyen de techniques de criblage à débit élevé, notamment la cytométrie en flux (FCM). Le procédé comprend les étapes consistant: 1) à développer des populations de cellules à cribler; 2) à colorer les populations de cellules au moyen d'un ou de plusieurs fluorochromes, de manière à produire une signature distincte d'excitation/émission pour chaque population de cellules; 3) à combiner des populations de cellules étiquetées en une suspension mélangée unique; 4) à analyser des populations aux fins de résolution de celles-ci en fonction de leur signature unique; et 5) à résoudre des populations individuelles et à effectuer une déconvolution des données afin d'extraire des informations significatives relatives aux populations.

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