C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 5/0793 (2010.01) C12N 5/071 (2010.01) C07K 14/33 (2006.01) C12N 5/10 (2006.01) C12Q 1/02 (2006.01) C12Q 1/37 (2006.01) G01N 33/02 (2006.01) G01N 33/68 (2006.01)
Patent
CA 2500040
The present invention provides a method of determining clostridial toxin activity by (a) contacting with a sample a cell containing a clostridial toxin substrate that includes a donor fluorophore; an acceptor having an absorbance spectrum overlapping the emission spectrum of the donor fluorophore; and a clostridial toxin recognition sequence containing a cleavage site that intervenes between the donor fluorophore and the acceptor, where resonance energy transfer is exhibited between the donor fluorophore and the acceptor under the appropriate conditions; (b) exciting the donor fluorophore; and (c) determining resonance energy transfer of the contacted cell relative to a control cell, where a difference in resonance energy transfer of the contacted cell as compared to the control cell is indicative of clostridial toxin activity.
L'invention concerne un procédé de détermination d'activité de toxine clostridienne consistant (a) à mettre en contact un échantillon de cellule contenant un substrat de toxine clostridienne comportant un fluorophore donneur, un accepteur dont le spectre d'absorption recouvre le spectre d'émission du fluorophore donneur et une séquence de reconnaissance de toxine clostridienne contenant un site de scission intervenant entre le fluorophore donneur et l'accepteur, le transfert d'énergie par résonance se produisant entre le fluorophore donneur et l'accepteur dans des conditions appropriées, (b) à exciter le fluorophore donneur, et (c) à déterminer l'énergie de transfert par résonance de la cellule en contact par rapport à une cellule témoin, une différence en transfert d'énergie de résonance entre la cellule en contact et la cellule témoin indiquant une activité de toxine clostridienne.
Aoki Kei Roger
Fernandez-Salas Ester
Steward Lance E.
Allergan Inc.
Gowling Lafleur Henderson Llp
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