C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/37 (2006.01) G01N 33/58 (2006.01) G01N 21/64 (2006.01)
Patent
CA 2581231
The present invention provides a method of determining the presence or activity of a clostridial toxin by (a) treating with a sample, under conditions suitable for clostridial toxin protease activity, a clostridial toxin substrate which includes a fluorophore; a bulking group; and a clostridial toxin recognition sequence containing a cleavage site that intervenes between the fluorophore and the bulking group; (b) exciting the fluorophore with plane polarized light; and (c) determining fluorescence polarization of the treated substrate relative to a control substrate, where a change in fluorescence polarization of the treated substrate as compared to fluorescence polarization of the control substrate is indicative of the presence or activity of the clostridial toxin.
La présente invention concerne une méthode qui permet de déterminer la présence ou l'activité d'une toxine clostridiale. La méthode consiste à: a) traiter au moyen d'un prélèvement, dans des conditions propres à déterminer l'activité protéase de la toxine clostridiale, un substrat de la toxine clostridiale comprenant un fluorophore, un groupe de mise en vrac, et une séquence d'identification de la toxine clostridiale contenant un site de clivage qui intervient entre le fluorophore et le groupe de mise en vrac; b) exciter le fluorophore avec une lumière polarisée plane; et c) déterminer la polarisation de fluorescence du substrat traité relativement à un substrat témoin, un changement de la polarisation de fluorescence du substrat traité par comparaison avec la polarisation de fluorescence du substrat témoin indiquant la présence ou l'activité de la toxine clostridiale.
Aoki Dei Roger
Gilmore Marcella A.
Steward Lance E.
Verhagen Marc
Williams Dudley J.
Allergan Inc.
Gowling Lafleur Henderson Llp
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