Enzymatic assays using umbelliferone substrates with cyclodextrins in droplets in oil
First Claim
1. A method of conducting an assay, the method comprising (a) incubating a droplet in oil, the droplet comprising an umbelliferone substrate, a sample potentially comprising an enzyme which cleaves the umbelliferone substrate, and a cyclodextrin compound;
- and (b) detecting a signal emitted from the droplet.
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Abstract
The invention is directed to droplet actuator devices and assay methods. The method may include, among other things, a method of conducting enzymatic assays using umbelliferone substrates with cyclodextrins in droplets in oil, the method including incubating a droplet in oil, the droplet preferably comprising an umbelliferone substrate, a sample, and a cyclodextrin compound. The methods may further include a method of substantially eliminating cross-contamination between droplets in enzymatic containing substrate-based bioassays. The method may include immobilization of the enzymatic substrate including forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the inclusion complex may be formed using cyclodextrins. In yet another embodiment, the invention may provide a method of enhancing hydrolysis of enzymatic substrates. The methods may further include a method of forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the substrate may be a 4-MU- or HMU containing substrate. The inclusion complex may be formed using cyclodextrin.
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Citations
7 Claims
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1. A method of conducting an assay, the method comprising (a) incubating a droplet in oil, the droplet comprising an umbelliferone substrate, a sample potentially comprising an enzyme which cleaves the umbelliferone substrate, and a cyclodextrin compound;
- and (b) detecting a signal emitted from the droplet.
- View Dependent Claims (2, 3, 4, 5, 6, 7)
Specification