Enzymatic substrates for multiple detection systems

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/34 (2006.01) C07C 237/22 (2006.01) C12Q 1/00 (2006.01) C07H 15/00 (2006.01) C07H 15/20 (2006.01)

Patent

CA 2718276

An inventive substrate is provided which includes a substrate compound of formula A-B1-B2-B3-B4: wherein A is a sugar moiety; B1 is a linker moiety allowing the conjugation of moiety A and the remaining structure of the substrate; B2 is a linker moiety with a free reactive amino group so as to be available for reaction with carboxylic acids or detectable tags; B3 contains a permanently charged element such as a quaternary ammonium group so as to increase sensitivity for mass spectrometry analysis; and B4 of various carbon length conferring specificity amongst individual substrates in detection methods. Also provided is a molecule of the formula B1-B2-B3-B4, with similar structural characteristics to an enzymatic product produced by the action of a target enzyme on an inventive substrate. Further provided are methods for using inventive substrates for detecting enzymatic activity.

Linvention concerne un substrat inventif qui comprend un composé de substrat de formule AB1B2B3B4 dans laquelle A est un fragment de sucre; B1 est un fragment de lieur permettant la conjugaison du fragment A et de la structure restante du substrat; B2 est un fragment de lieur avec un groupe amino réactif libre de façon à ce quil disponible pour une réaction avec des acides carboxyliques ou des marqueurs détectables; B3 contient un élément chargé de façon permanente tel quun groupe ammonium quaternaire de façon à augmenter la sensibilité à lanalyse par spectrométrie de masse; et B4 de longueur datome de carbone diverse conférant une spécificité parmi les substrats individuels dans des procédés de détection. Il est également proposé une molécule de formule B1B2B3B4, avec des caractéristiques structurelles similaires à un produit enzymatique produit par laction dune enzyme cible sur un substrat inventif. Il est également proposé des procédés dutilisation de substrats inventifs pour détecter une activité enzymatique.

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