Enzyme cycling based assays for alpha-methylacyl-coa racemase

Details Enzyme cycling based assays for alpha-methylacyl-coa racemase Enzyme cycling based assays for alpha-methylacyl-coa racemase

C - Chemistry, Metallurgy

12

Q

C12Q 1/26 (2006.01) C12Q 1/00 (2006.01) G01N 33/53 (2006.01)

Patent

CA 2583546

The present invention provides a method for assaying alpha-methylacyl-CoA racemase activity. In the assay, a sample containing an alpha-methylacyl-CoA racemase or suspected of containing an alpha-methylacyl-CoA racemase is contacted with (2R)-2-methylacyl-CoA. If alpha-methylacyl-CoA racemase is present in the sample (2R)-2-methylacyl-CoA is converted into (2S)-methylacyl- CoA. The method then utilizes a cycling reaction system between (2S)- methylacyl-CoA and trans-2,3-dehydroacyl-CoA to generate a detectable signal that corresponds to the alpha-methylacyl-CoA racemase activity. Kits for assaying alpha-methylacyl-CoA racemase based on the same principle are also provided.

L'invention concerne un procédé pour doser l'activité de la racémase alpha-méthylacyl-CoA. Dans l'essai biologique, un échantillon contenant une racémase alpha-méthylacyl-CoA ou suspectée de contenir une racémase alpha-méthylacyl-CoA est mise en contact avec du (2R)-2-méthylacyle-CoA. Si la racémase alpha-méthylacyl-CoA est présente dans l'échantillon, le (2R)-2-méthylacyl-CoA est converti en (2S)-2-méthylacyl-CoA. Ledit procédé comprend ensuite l'utilisation d'un système de réaction en cycle entre le (2S)-2-méthylacyl-CoA et le trans-2,3-déshydroacyl-CoA, de manière à produire un signal, pouvant être détecté, qui correspond à l'activité de la racémase alpha-méthylacyl-CoA. L'invention concerne également des kits destinés à doser la racémase alpha-méthylacyl-CoA fondés sur le même principe.

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