Multisite biosensor and associated method

• US 9,927,435 B2
• Filed: 10/15/2009
• Issued: 03/27/2018
• Est. Priority Date: 10/15/2009
• Status: Active Grant

• Current Assignee: Robert Bosch GmbH, Stanford University
• Original Assignee: Robert Bosch GmbH, Stanford University
• Inventors: Kavusi, Sam, Roser, Daniel, Lang, Christoph, Talasaz, AmirAli Haj Hossein
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US12/580,113
• Publication Number:
  US 20110091870A1
• Time in Patent Office: 3,085 Days
• Field of Search: 73826046, 356478, 4352883, 4352884, 435 71, 436174-180, 436501, 436518, 436523-536, 436809
• CPC Class Codes: G01N 29/32 Arrangements for suppressing undesired influences, e.g. temperature or pressure variations , compensating for signal noise
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

A method of detecting a biomarker in one embodiment includes identifying a quantity of biomolecule types in a sample, exposing the sample to a plurality of test sites, wherein the number of test sites in the plurality of test sites is equal to or greater than the identified quantity of biomolecule types, establishing, for each of the plurality of test sites, a respective test environment, wherein the test environment for each of the plurality of test sites is different from the test environment for each of the other of the plurality of test sites, obtaining a detection signal associated with each of the plurality of test sites, and determining the concentration of one of the biomolecule types based upon the obtained detection signals.

Claims

1. A method of detecting a biomarker comprising:

identifying a number of biomolecule types in a sample with an unknown concentration of the biomolecule types and at least two biomolecule types;

exposing the sample to a plurality of test sites, wherein the number of test sites in the plurality of test sites is based upon the identified number of biomolecule types so as to be equal to or greater than the identified number of biomolecule types;

establishing, for each of the plurality of test sites, a respective test environment, wherein the test environment for each of the plurality of test sites is different from the test environment for each of the other of the plurality of test sites, and each of the plurality of test sites is configured to capture each of the biomolecule types;

identifying, for each of the identified biomolecule types, a binding efficiency for each of the test environments for each of the plurality of test sites;

obtaining a detection signal associated with each of the plurality of test sites; and

calculating the concentration of one of the biomolecule types based upon the obtained detection signals and the identified binding efficiencies.

2. The method of claim 1, wherein calculating the concentration comprises:

applying at least one linear estimation algorithm to the obtained detection signals.

3. The method of claim 1, wherein establishing a respective test environment comprises:

controlling at least one environmental factor of a group of environmental factors consisting of temperature, electric field, magnetic field, pH, and buffer type for each of the plurality of test sites, such that the controlled at least one environmental factor is different between at least two of the plurality of test sites.

4. The method of claim 3, wherein controlling at least one environmental factor comprises:

controlling the at least one environmental factor such that the controlled at least one environmental factor at each of the plurality of sites is different from each of the other of the plurality of test sites.

5. The method of claim 1, wherein establishing a respective test environment comprises:

providing at least one capturing agent;

identifying, for each of the plurality of test sites, a respective capturing agent recipe, wherein the capturing agent recipe for each of the plurality of test sites is different from the capturing agent recipe for each of the other of the plurality of test sites; and

immobilizing, in each of the plurality of test sites, the at least one capturing agent in accordance with the identified recipe.

6. The method of claim 5, wherein the at least one capturing agent comprises a plurality of capturing agents, each of the plurality of capturing agents exhibiting an affinity to one or more of the identified biomolecule types that is different from the affinity to the one or more of the identified biomolecule types of another of the plurality of capturing agents.

7. The method of claim 6, wherein the capturing agent recipe for each of the plurality of test sites includes a selected one of the at least one capturing agent that is different from the at least one capturing agent in the other of the plurality of test sites.

8. The method of claim 5, wherein the capturing agent recipe for a first of the plurality of test sites includes a selected one of the at least one capturing agent that is the same as the at least one capturing agent in a second of the plurality of test sites.

9. The method of claim 8, wherein the concentration of the selected one of the at least one capturing agent in the first of the plurality of test sites is greater than the concentration of the selected one of the at least one capturing agent in the second of the plurality of test sites.

10. The method of claim 1, further comprising:

forming each of the plurality of test sites in a respective well of a multi-well plate.

11. The method of claim 1, further comprising:

forming each of the plurality of test sites on a CMOS substrate.

17. The method of claim 2, wherein calculating the concentration of one of the biomolecule types based upon the obtained detection signals comprises:

calculating the concentration of one of the biomolecule types based upon the following equations;

S₁=α

_1-1C₁+α

_1-2C₂

S₂=α

_2-1C₁+α

_2-2C₂

wherein

S₁ is a first obtained signal associated with a first test site of the plurality of test sites,

S₂ is a second obtained signal associated with a second test site of the plurality of test sites,

α

_1-1 is a binding efficiency proportional to an affinity for a first of the identified at least two biomolecule types at the environment established in the first test site,

α

_1-2 is a binding efficiency proportional to an affinity for a second of the identified at least two biomolecule types at the environment established in the first test site,

α

_2-1 is a binding efficiency proportional to an affinity for the first of the identified at least two biomolecule types at the environment established in the second test site,

α

_2-2 is a binding efficiency proportional to an affinity for the second of the identified at least two biomolecule types at the environment established in the second test site,

C₁ is a concentration in the sample of the first of the identified at least two biomolecule types, and

C₂ is a concentration in the sample of the second of the identified at least two biomolecule types.

18. The method of claim 1, wherein calculating the concentration of one of the biomolecule types based upon the obtained detection signals does not include:

calculating a respective concentration with each of the obtained detection signals.

12. A method of determining a concentration of a biomarker in a sample comprising:

identifying a number of detection signal contributors for a plurality of detection signals, wherein at least one of the detection signal contributors is a molecule of interest;

exposing a sample with an unknown concentration of the molecule of interest to a plurality of test sites, wherein the number of test sites in the plurality of test sites is based upon the identified number of detection signal contributors so as to be equal to or greater than the identified number of detection signal contributors and each of the plurality of test sites is configured to capture each of the detection signal contributors;

establishing, for each of the plurality of test sites, a respective test environment, wherein the test environment for each of the plurality of test sites is different from the test environment for each of the other of the plurality of test sites;

identifying, for each of the detection signal contributors, a binding efficiency for each of the test environments for each of the plurality of test sites;

obtaining a respective one of the plurality of detection signals from each of the plurality of test sites; and

calculating the concentration of the molecule of interest based upon the obtained plurality of detection signals and the identified binding efficiencies.

13. The method of claim 12, wherein establishing a respective test environment comprises:

controlling at least one environmental factor of a group of environmental factors consisting of temperature, electric field, magnetic field, and pH, for each of the plurality of test sites, such that the controlled at least one environmental factor is different between at least two of the plurality of test sites.

14. The method of claim 13, wherein controlling at least one environmental factor comprises:

controlling the at least one environmental factor such that the controlled at least one environmental factor at each of the plurality of sites is different from each of the other of the plurality of test sites.

15. The method of claim 12, wherein establishing a respective test environment comprises:

providing at least one capturing agent;

identifying, for each of the plurality of test sites, a respective capturing agent recipe, wherein the capturing agent recipe for each of the plurality of test sites is different from the capturing agent recipe for each of the other of the plurality of test sites; and

immobilizing, in each of the plurality of test sites, the at least one capturing agent in accordance with the identified recipe.

16. The method of claim 15, wherein the at least one capturing agent comprises a plurality of capturing agents, each of the plurality of capturing agents exhibiting an affinity to one or more of the identified biomolecule types that is different from the affinity to the one or more of the identified biomolecule types of another of the plurality of capturing agents.