Lipophilic dye-based fret assays for clostridial toxin activity

G - Physics – 01 – N

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G01N 33/50 (2006.01)

Patent

CA 2604039

Compositions useful for detecting Clostridial toxin activity comprising a cell that comprises a membrane-associated Clostridial toxin substrate comprising a first member of a fluorescence resonance energy transfer pair; and a Clostridial toxin recognition sequence including a cleavage site; and a membrane-associated second member of the FRET pair and methods useful for determining Clostridial toxin activity using such Clostridial toxin substrates.

L'invention porte sur des compositions utilisées pour détecter l'activité de la toxine clostridiale, lesquelles compositions comprennent une cellule qui renferme: un substrat de toxine clostridiale associé à une membrane, comportant un premier élément d'une paire de transfert d'énergie fluorescente de résonance ("fluorescence resonance energy transfer" ou FRET) et une séquence de reconnaissance de la toxine clostridienne comprenant un site de clivage; et un second élément d'une paire FRET associé à une membrane. L'invention concerne également des procédés qui permettent de déterminer l'activité de la toxine clostridiale au moyen desdits substrats de toxine clostridiale.

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