High throughput assays for the proteolytic activities of...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/37 (2006.01) C07K 7/08 (2006.01) C07K 14/33 (2006.01) C07K 17/00 (2006.01) G01N 33/542 (2006.01) G01N 33/543 (2006.01) G01N 33/569 (2006.01)

Patent

CA 2451909

In this application is described substrates for high-throughput assays of clostridial neurotoxin proteolytic activities. Two types of substrates are described for use in assays for the proteolytic activities of clostridial neurotoxins: (1) modified peptides or proteins that can serve as FRET substrates and (2) modified peptides or proteins that can serve as immobilized substrates. In both types a fluorescent molecules is present in the substrate, eliminating the requirement for the addition of a fluorigenic reagent. The assays described can be readily adapted for use in automated or robotic systems.

L'invention concerne des substrats destinés à des tests de détection rapide des activités protéolytiques des neurotoxine clostridiales. Cette invention comprend deux types de substrats destinés à des tests permettant de détecter les activités protéolytiques des neurotoxines clostridiales : (1) des peptides ou des protéines modifiés pouvant servir de substrats pour un procédé FRET (transfert d'énergie par résonance de fluorescence) et (2) des peptides ou des protéines modifiés pouvant servir de substrats immobilisés. Dans ces deux types de substrats, une molécule fluorescente est présente dans le substrat de sorte qu'il n'est pas nécessaire d'ajouter un réactif fluorogène. On peut facilement adapter ces systèmes d'analyse de manière à pouvoir les utiliser dans des systèmes automatisés ou robotiques.

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