Method of conducting a droplet based enzymatic assay
First Claim
1. A method of conducting a droplet-based enzymatic assay, the method comprising:
- (a) providing a droplet actuator;
(b) providing on the droplet actuator;
(i) a droplet comprising an enzyme of interest;
(ii) a droplet comprising a substrate which is potentially modified in the presence of the enzyme;
(c) executing droplet operations on the droplet actuator to combine the droplet of (b)(i) with the droplet of (b)(ii), thereby yielding an assay droplet, wherein the droplet operations are electrowetting mediated;
(d) detecting modification of the substrate by the enzyme in the assay droplet on the droplet actuator; and
wherein each of the droplet of (b)(i), the droplet of (b)(ii) and the assay droplet is at least partially surrounded by a silicone oil filler fluid comprising an nonionic low hydrophile-lipophile balanced (HLB) surfactant.
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Abstract
A method of conducting a droplet-based enzymatic assay is provided. The method generally makes use of a droplet actuator. A droplet comprising an enzyme of interest is provided on the droplet actuator along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. The method involves executing droplet operations on the droplet actuator to combine the droplets, thereby yielding an assay droplet, and detecting modification of the substrate by the enzyme in the assay droplet on the droplet actuator. The enzyme of interest may, for example, be a potentially mutated or improperly folded enzyme exhibiting altered enzyme activity as compared to a corresponding normal enzyme.
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Citations
39 Claims
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1. A method of conducting a droplet-based enzymatic assay, the method comprising:
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(a) providing a droplet actuator; (b) providing on the droplet actuator; (i) a droplet comprising an enzyme of interest; (ii) a droplet comprising a substrate which is potentially modified in the presence of the enzyme; (c) executing droplet operations on the droplet actuator to combine the droplet of (b)(i) with the droplet of (b)(ii), thereby yielding an assay droplet, wherein the droplet operations are electrowetting mediated; (d) detecting modification of the substrate by the enzyme in the assay droplet on the droplet actuator; and wherein each of the droplet of (b)(i), the droplet of (b)(ii) and the assay droplet is at least partially surrounded by a silicone oil filler fluid comprising an nonionic low hydrophile-lipophile balanced (HLB) surfactant. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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Specification