Methods for determining the concentration of an analyte in solution

Patent metadata

Field	Value
Publication number	US-9809838-B2
Application number	US-201615290939-A
Country	US
Kind code	B2
Filing date	Oct 11, 2016
Priority date	Aug 30, 2007
Publication date	Nov 7, 2017
Grant date	Nov 7, 2017

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.

First claim

The invention claimed is:

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.

Assignees

• Tufts College

Inventors

• Walt David R
• Rissin David M

Classifications

• C12Q1/40 Primary

  involving amylase · CPC title

• G01N33/6854

  Immunoglobulins · CPC title

• G01N2333/522

  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 title

• G01N2333/938

  acting on beta-galactose-glycoside bonds, e.g. beta-galactosidase · CPC title

• G01N33/54366 Primary

  Apparatus specially adapted for solid-phase testing · CPC title