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 microfluidic device for detection of an active enzymatic agent, the device comprising:
a substrate defining at least a portion of a first channel, a second channel, and a third channel coupled between the first channel and the second channel; a first membrane positioned in the first channel and configured to block a substrate molecule including a reporter moiety and a sequence configured to be cleaved by the active enzymatic agent; at least one reagent loading channel coupled to the first channel, the third channel coupled to the first channel at a location between the reagent loading channel and the first membrane; a second membrane positioned in the second channel and configured to concentrate reaction products generated following exposure of the active enzymatic agent to the substrate molecule; and wherein the third channel is configured to transport reaction products from a vicinity of the first membrane to a vicinity of the second membrane; and a separation channel including a separation gel, the separation channel coupled to the second channel and configured for separation of the reaction products, an elution time of the reporter moiety to a detection region in the separation channel being indicative of presence or absence of the active enzymatic agent in the sample.