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US 11,187,702 B2Granted
Enzyme quantification
AssigneeBio-Rad Laboratories, Inc.
InventorsDarren R. Link, Michael L. Samuels
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Abstract
The invention generally relates to methods for quantifying an amount of enzyme molecules. Systems and methods of the invention are provided for measuring an amount of target by forming a plurality of fluid partitions, a subset of which include the target, performing an enzyme-catalyzed reaction in the subset, and detecting the number of partitions in the subset. The amount of target can be determined based on the detected number.
Claims10
A method of performing a digital ELISA assay in droplets, the method comprising:
- providing reagent droplets comprising reporter antibodies and second antibodies bound to magnetic beads into a channel of a microfluidic device;
- introducing test droplets comprising a test fluid into the microfluidic device;
- combining the reagent droplets with the test droplets in the microfluidic device to form ELISA sandwiches, each ELISA sandwich comprising one bead-bound antibody, a target analyte from the test fluid, and one reporter antibody;
- adding, via the microfluidic device, a fluorogenic substrate into droplets that contain one of the ELISA sandwiches;
- and
- detecting fluorescence indicating presence of the target analyte, wherein the detecting step provides a digital count of a number of molecules of analyte per volume of the test fluid.
3 more elements
Dependencies are inferred from the claim text.