BOTULINUM TOXIN ASSAY WITH IMPROVED SENSITIVITY

摘要

Methods and compositions are provided where a transfected cell that produces a hybrid protein with a reporter-containing portion and a botulinum toxin cleavage site is contacted with a botulinum toxin at elevated temperatures and/or in media having a reduced sodium concentration. Kits that include such media and a botulinum toxin are also described.

主权项

1. A method of increasing the sensitivity of cell-based detection of a botulinum toxin, comprising:
(i) providing a transfected cell that produces a construct comprising;
(a) a first terminus comprising a reporter-containing portion, wherein the reporter-containing portion exhibits a signal; and,(b) a cleavage site that interacts with the botulinum toxin in a manner that produces a cleavage of the reporter-containing portion from a remainder of the construct; (ii) exposing the transfected cell to the botulinum toxin at a toxin exposure temperature of from 38° C. to 41° C., wherein sensitivity of the transfected cell's response to the Botulinum toxin at the toxin exposure temperature increases at least 2 fold compared to the transfected cell's sensitivity to the botulinum toxin at 37° C.; (iv) obtaining the signal from the reporter-containing portion.