Enzyme quantification
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 sunset. The amount of target can be determined based on the detected number.
1. A method for identifying components of a chemical reaction, the method comprising:
forming fluid partitions comprising components of a chemical reaction, wherein said components comprise: an enzyme that catalyzes said chemical reaction, and a detectable label that is acted on by said chemical reaction;
conducting said chemical reaction; and
determining an amount of said enzyme in said fluid partitions based upon one or more properties of the detectable label in said fluid partitions.
2. The method of claim 1 , wherein the one or more properties of the detectable label includes amount of the detectable label.
3. The method of claim 2 , wherein the amount of the detectable label is detected by an optical property.
4. The method of claim 1 , further comprising the step of identifying fluid partitions that contain a released detectable label.
5. The method of claim 1 , wherein said components comprise an enzyme and at least one substrate of the enzyme.
6. The method of claim 5 , wherein said enzyme catalyzes a reaction that results in release of the detectable label from said substrate.
7. The method of claim 1 , further comprising determining a number of enzyme molecules within each partition based upon signal strength of the detectable label.
8. The method of claim 1 , wherein determining the amount of the least one of said components is based upon a localized concentration of the detectable label.
9. The method of claim 8 , wherein the localized concentration is detected within a fluid partition.
10. The method of claim 1 , wherein said fluid partitions are droplets.
11. The method of claim 10 , wherein the droplets are surrounded by an immiscible carrier fluid.
12. The method of claim 1 , wherein determining the amount of the enzyme is based upon a ratio of a localized increase to partition wide decrease in signal intensity.