METHODS, MICROFLUIDIC DEVICES, AND SYSTEMS FOR DETECTION OF AN ACTIVE ENZYMATIC AGENT

摘要

Embodiments of the present invention provide methods, microfluidic devices, and systems for the detection of an active target agent in a fluid sample. A substrate molecule is used that contains a sequence which may cleave in the presence of an active target agent. A SNAP25 sequence is described, for example, that may be cleaved in the presence of Botulinum Neurotoxin. The substrate molecule includes a reporter moiety. The substrate molecule is exposed to the sample, and resulting reaction products separated using electrophoretic separation. The elution time of the reporter moiety may be utilized to identify the presence or absence of the active target agent.

主权项

1. A method for detecting enzymatic activity of an active target agent in a sample, the method comprising:
concentrating said sample at a membrane within a microfluidic device; concentrating a substrate molecule larger than 25 kDa at the membrane within the microfluidic device, the substrate molecule comprising a reporter moiety and a protein sequence configured to cleave in the presence of the active target agent into reaction products, at least one reaction product including the reporter moiety and having a size larger than 25 kDa; measuring an elution time for the reporter moiety to determine cleavage of said sequence, said cleavage at least in part identifying the presence of the active target agent in said sample.