Microfluidic in-line labeling method of continuous-flow protease inhibition analysis
AssigneeCaliper Technologies Corp.
InventorsHua Yang, Steven Sundberg
Life of this patent
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Abstract
Enzyme assays are performed in microfluidic devices including, e.g., in-line labeling, separation, and detection of assay products. In-line labeling allows assays, e.g., protease assays, to be performed in a continuous flow microfluidic format. Also included are microfluidic devices and integrated systems for performing in-line labeling in continuous flow enzyme assays.
Claims30
A method for screening a plurality of compounds for modulators that affect the activity of a protease, the method comprising:
- (i) providing a plurality of compounds;
- (ii) introducing a compound selected from the plurality of compounds into a microscale channel;
- (iii) contacting the compound with a protease and a protease substrate in the microscale channel, thereby reacting the protease substrate with the protease in the presence of the compound to produce one or more products;
- (iv) labeling the one or more products by flowing a labeling reagent into the at least one microscale cavity or into the additional microscale cavity to react with the one or more products and produce one or more labeled products;
- (v) detecting the amount of one or more of the labeled products to determine the level of inhibition or activation of the compound;
- and
- (vi) repeating steps (ii) through (v) for each of the plurality of compounds.
A method for determining the inhibition or activation rate constants for a modulator that affects the activity of a protease, the method comprising:
- (i) providing a modulator;
- (ii) introducing the modulator into a microscale channel;
- (iii) contacting the modulator with a protease and a protease substrate in the microscale channel, thereby reacting the protease substrate with the protease in the presence of the modulator to produce one or more products;
- (v) labeling the one or more products by flowing a labeling reagent into the at least one microscale cavity or into the additional microscale cavity to react with the one or more products and produce one or more labeled products;
- and
- (vi) detecting the amount of one or more of the labeled products to determine the level of inhibition or activation of the compound;
- and
- (vii) determining the rate constant for the modulator.
Dependencies are inferred from the claim text.