Longitudinal assay

• US 10,060,919 B2
• Filed: 01/22/2015
• Issued: 08/28/2018
• Est. Priority Date: 11/24/2010
• Status: Active Grant

• Current Assignee: Inanovate, Inc.
• Original Assignee: Inanovate, Inc.
• Inventors: Nelson, James Curtis, Ure, David
• Primary Examiner(s): COUNTS, GARY W
• Application Number:
  US14/603,329
• Publication Number:
  US 20150141288A1
• Time in Patent Office: 1,314 Days
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: F04B 43/0009 Special features
• CPC Class Codes: F04B 43/02 having plate-like flexible members, e.g. diaphragms F04B43/14 takes precedence
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/557 using kinetic measurement, i.e. time rate of progress of an antigen-antibody interaction

Abstract

The invention relates to macro and small molecule detection and, more particularly, to methods for detecting macro and small molecules, including bio-molecules, in a liquid or gaseous sample.

Claims

1. A method of distinguishing specific binding of an analyte and capture agent from non-specific binding of non-analyte molecules to the capture agent, the method comprising:

constructing a binding curve for the binding of the capture agent to an analyte, a non-analyte, or both in a sample based on a plurality of temporally-spaced detections of the capture agent within a single assay;

detecting a first detection intensity of the binding of the capture agent and a first temporal point in the binding curve at which a rise to an equilibrium has been reached or is inferred; and

if a rise to equilibrium has been reached, attributing the first detection intensity to non-specific binding;

or

if a rise to equilibrium is inferred, attributing to non-specific binding a first portion of the binding curve subsequent to the first temporal point that is equal to the first detection intensity.

2. The method of claim 1, further comprising:

in the case that a rise to equilibrium is inferred, attributing to specific binding a second portion of at least one additional detected intensity subsequent to the first temporal point equal to a difference between the at least one additional detected intensity and the first detection intensity.

7. The method of claim 1, wherein, in the case that a rise to equilibrium is inferred, the method further comprises:

attributing to specific binding of the analyte a second portion of the binding curve at an additional detected intensity subsequent to the first temporal point equal to a difference between the additional detected intensity and the first detection intensity; and

determining an initial concentration of the analyte based on the specific binding.

3. A method of distinguishing specific binding of an analyte and capture agent from non-specific binding of non-analyte molecules to the capture agent, the method comprising:

constructing a binding curve for the binding of the capture agent to an analyte, a non-analyte, or both in a sample based on a plurality of temporally-spaced detections of the capture agent within a single assay;

detecting a first detection intensity of the binding of the capture agent and a first temporal point in the binding curve at which a rise to an equilibrium has been reached; and

attributing the first detection intensity to non-specific binding.

4. The method of claim 3, further comprising:

attributing detection intensities occurring subsequent to the first temporal point to non-specific binding; and

concluding that the detection intensities of the binding curve do not include a specific binding component.

5. A method of identifying biomarkers for therapeutic and/or diagnostic use, the method comprising:

constructing a binding curve for the binding of a capture agent to an analyte, a non-analyte, or both in a sample based on a plurality of temporally-spaced detections of the capture agent within a single assay;

detecting a first detection intensity of the capture agent and a first temporal point in the binding curve at which a rise to an equilibrium is inferred;

attributing to non-specific binding a first portion of the binding curve at an additional detected intensity subsequent to the first temporal point that is equal in intensity to the first detection intensity;

attributing to specific binding of the analyte a second portion of the binding curve at the additional detected intensity subsequent to the first temporal point equal to a difference between the additional detected intensity and the first detection intensity; and

determining an initial concentration of the analyte based on the specific binding.

6. The method of claim 5, further comprising:

in the case that the initial concentration of the analyte is determined to be less than about 1% of the sample, identifying the capture agent to analyte binding as a potential biomarker for therapeutic and/or diagnostic use.