Detecting and profiling molecular complexes

Details Detecting and profiling molecular complexes Detecting and profiling molecular complexes

C - Chemistry, Metallurgy

07

K

C07K 16/46 (2006.01) C12N 7/02 (2006.01) G01N 33/53 (2006.01)

Patent

CA 2535510

Methods are provided for detecting the formation of complexes of molecules, especially proteins, in a sample, such as a cell or tissue lysate. In one aspect, a cleaving probe specific for a first protein in a complex and one or more binding compounds specific for one or more second proteins in a complex are provided. Upon binding, the cleaving probe is induced to generate an active species, such as singlet oxygen, that cleaves molecular tags attached to the binding compounds only in the local region of the cleaving probe. The released molecular tags are separated from the assay mixture and from one another to provide a readout that is related to the number and types of proteins present in the complex.

L'invention concerne des procédés permettant de détecter la formation de complexes de molécules, notamment de protéines, dans un échantillon tel qu'un lysat de cellules ou de tissu. Dans un aspect, elle concerne une sonde de clivage spécifique d'une première protéine d'un complexe, et un ou plusieurs composés de liaison spécifiques d'une ou de plusieurs secondes protéines d'un complexe. Lors de la liaison, la sonde de clivage produit une espèce active, telle que l'oxygène singulet, qui coupe les marqueurs moléculaires fixés aux composés de liaison, uniquement dans la région locale de la sonde de clivage. Les marqueurs moléculaires libérés sont séparés les uns des autres ainsi que du mélange analysé pour former un résultat qui est lié au nombre et aux types de protéines présentes dans le complexe.

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