Please download the dossier by clicking on the dossier button x
×

Selection by compartmentalised screening

  • US 9,448,172 B2
  • Filed: 09/29/2005
  • Issued: 09/20/2016
  • Est. Priority Date: 03/31/2003
  • Status: Active Grant
First Claim
Patent Images

1. A method for screening compounds, the method comprising:

  • introducing a first aqueous fluid comprising a target enzyme and a second aqueous fluid comprising a repertoire of small molecule compounds by separately injecting the first aqueous fluid and the second aqueous fluid into an immiscible fluorocarbon oil continuously flowing through a first microfluidic channel to form a plurality of first water-in-fluorocarbon oil microcapsules, wherein the first immiscible fluorocarbon oil comprises a fluorosurfactant, and wherein each aqueous portion of each of the plurality of the first water-in-fluorocarbon oil microcapsule comprises the target enzyme and a subset of the repertoire of small molecule compounds;

    continuously flowing the plurality of the first water-in-fluorocarbon oil microcapsules through the first microfluidic channel and conducting a first target enzyme catalyzed reaction between the target enzyme and the subset of the repertoire of small molecule compounds within the aqueous portion of each of the plurality of the first water-in-fluorocarbon oil microcapsules to produce a first reaction product;

    introducing a third aqueous fluid comprising a fluorogenic substrate from a second microfluidic channel, wherein the fluorogenic substrate is specific for the target enzyme;

    merging a plurality of second water-in-fluorocarbon oil microcapsules flowing through the second microfluidic channel with the plurality of the first water-in-fluorocarbon oil microcapsules flowing through the first microfluidic channel to form a plurality of merged water-in-fluorocarbon oil microcapsules, wherein the aqueous portion of each of the plurality of second water-in-fluorocarbon oil microcapsules comprises the fluorogenic substrate, to form a plurality of merged water-in-fluorocarbon oil microcapsules, wherein the aqueous portion of the plurality of merged water-in-fluorocarbon oil microcapsules comprises the first reaction product and the fluorogenic substrate;

    conducting a second enzyme catalyzed reaction between the first reaction product and the fluorogenic substrate within the aqueous portion of each of the plurality of the merged water-in-fluorocarbon oil microcapsules to produce a second reaction product; and

    optically detecting the second reaction product.

View all claims
  • 3 Assignments
Timeline View
Assignment View
    ×
    ×