Methods for determining the concentration of an analyte in solution

• US 9,809,838 B2
• Filed: 10/11/2016
• Issued: 11/07/2017
• Est. Priority Date: 08/30/2007
• Status: Active Grant

• Current Assignee: Trustees of Tufts College
• Original Assignee: Trustees of Tufts College
• Inventors: Walt, David R., Rissin, David M.
• Primary Examiner(s): Gross, Christopher M
• Application Number:
  US15/290,939
• Publication Number:
  US 20170159104A1
• Time in Patent Office: 392 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/40 involving amylase
• CPC Class Codes: G01N 2333/522 Alpha-chemokines, e.g. NAP-2, ENA-78, GRO-alpha/MGSA/NAP-3, GRO-beta/MIP-2alpha, GRO-gamma/MIP-2beta, IP-10, GCP-2, MIG, PBSF, PF-4 or KC
• CPC Class Codes: G01N 2333/938 acting on beta-galactose-glycoside bonds, e.g. beta-galactosidase
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

Disclosed is a method for measuring the concentration of an analyte or analytes in a solution. Although the methods can be conducted using a number of different assay formats, in one embodiment, the assays are conducted in reaction vessels defined, at least in part, by the distal ends of fiber optic strands.

Claims

1. A method of determining the concentration of an analyte in a fluid sample to be tested, the method comprising the steps of:

(a) partitioning at least a portion of the analyte molecules in the fluid sample across a plurality of reaction vessels so that more than 20% but less than 95% of the reaction vessels contain at least one molecule, wherein the analyte is attached to a nanoparticle or microparticle by a capture component;

(b) determining the presence or absence of the analyte in each reaction vessel to identify the number of reaction vessels that contain analyte and/or to identify the number of reaction vessels that contain no analyte; and

(c) determining the concentration of the analyte in the fluid sample by a Gaussian distribution analysis of the number of reaction vessels that contain the analyte.

2. The method of claim 1, wherein, in step (a), more than 20% but less than 60% of the reaction vessels contain at least one molecule.

3. The method of claim 1, wherein, in step (a), more than 60% but less than 95% of the reaction vessels contain at least one molecule.

4. The method of claim 1, wherein, in step (a), the sample to be tested is partitioned into at least 1,000 reaction vessels.

5. The method of claim 1, wherein, in step (a), the sample to be tested is partitioned into from 10,000 to 200,000 reaction vessels.

6. The method of claim 5, wherein the sample to be tested is partitioned into from 50,000 to 100,000 reaction vessels.

7. The method of claim 1, wherein the analyte is a biomolecule.

8. The method of claim 7, wherein the biomolecule is selected from the group consisting of a protein, a nucleic acid, a lipid, and a carbohydrate.

9. The method of claim 1, wherein the reaction vessels have a volume of from 10 attoliters to 50 picoliters.

10. The method of claim 1, wherein at least a portion of the reaction vessel is defined by a distal end of an optical fiber.