Method of conducting an assay

• US 8,592,217 B2
• Filed: 01/10/2013
• Issued: 11/26/2013
• Est. Priority Date: 03/22/2007
• Status: Active Grant

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Eckhardt, Allen E.
• Primary Examiner(s): COLE, MONIQUE T
• Application Number:
  US13/738,259
• Publication Number:
  US 20130130936A1
• Time in Patent Office: 320 Days
• Field of Search: 436/86
• US Class Current: 436/86
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: C07D 311/16 substituted in position 7
• CPC Class Codes: C07D 311/18 substituted otherwise than in position 3 or 7
• CPC Class Codes: C07D 311/20 hydrogenated in the hetero ring
• CPC Class Codes: C07D 405/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07D 413/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07H 15/203 Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
• CPC Class Codes: C07H 17/075 Benzo[b]pyran-2-ones
• CPC Class Codes: C12Q 1/25 involving enzymes not classifiable in groups C12Q1/26 - C12Q1/66
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/40 involving amylase
• CPC Class Codes: C12Q 1/44 involving esterase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Y 302/01023 Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 2333/916 : acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
• CPC Class Codes: G01N 2333/926 : acting on alpha -1, 4-glucosidic bonds, e.g. hyaluronidase, invertase, amylase
• CPC Class Codes: G01N 2333/938 : acting on beta-galactose-glycoside bonds, e.g. beta-galactosidase
• CPC Class Codes: G01N 2800/04 : Endocrine or metabolic disorders
• CPC Class Codes: G01N 33/5438 : Electrodes

Abstract

The invention is directed to droplet actuator devices and assay methods. The invention includes assay methods of conducting an assay comprising combining a sample with an umbelliferyl derivative, wherein the sample potentially comprises an enzyme capable of cleaving the umbelliferyl derivative and where the umbelliferyl derivative comprises an umbelliferyl core modified with one or more modifying moieties.

Claims

1. A method of conducting an assay comprising:

(I) combining a sample droplet with a droplet comprising an umbelliferyl derivative to yield an assay droplet wherein;

(a) the sample potentially comprises an enzyme capable of cleaving the umbelliferyl derivative; and

(b) the umbelliferyl derivative is a modified version of an umbelliferyl parent compound comprising an umbelliferyl core modified with one or more modifying moieties, said one or more modifying moieties;

(i) selected to increase hydrophilicity of the derivative relative to the parent compound without eliminating the fluorescence of the umbelliferyl core upon cleavage by the enzyme; and

(ii) coupled to the umbelliferyl core at a site that remains with the umbelliferyl core following cleavage by the compound; and

(II) detecting a signal from the assay droplet which is indicative of the presence of an enzyme capable of cleaving the umbelliferyl derivative.

2. The method of claim 1, wherein the droplets are in oil.

3. The method of claim 1, wherein the droplets are surrounded by oil.

4. The method of claim 1, wherein the combining is conducted using droplet operations mediated by electrodes on a droplet actuator.

5. The method of claim 1, wherein the umbelliferyl derivatives comprise an umbelliferyl core coupled to a hydrophilic polymer or oligomer group.

6. The method of claim 5 wherein the hydrophilic polymer or oligomer group comprises a salt moiety.

7. The method of claim 1, wherein the umbelliferyl derivatives comprise an umbelliferyl core coupled to a moiety comprising a polyalkalene glycol group.

8. The method of any of claim 1 and following, wherein the umbelliferyl derivatives comprise an umbelliferyl core coupled to a moiety comprising a polyethylene glycol group.

9. The method of claim 1, wherein the umbelliferyl derivatives comprise an umbelliferyl core coupled to a moiety comprising a polypropylene glycol group.

10. The method of claim 1, wherein the umbelliferyl derivatives comprise an umbelliferyl core coupled to a sulfonic acid.

11. The method of claim 1, wherein the umbelliferyl derivatives comprise an umbelliferyl core coupled to an amino group.