Efficient sensor for detecting and determining the concentrations of targets

• US 9,683,258 B2
• Filed: 05/14/2010
• Issued: 06/20/2017
• Est. Priority Date: 05/14/2010
• Status: Expired due to Fees

• Current Assignee: Sharp Kabushiki Kaisha (Hon Hai Precision Industry Co., Ltd.)
• Original Assignee: Sharp Laboratories of America Incorporated (Hon Hai Precision Industry Co., Ltd.)
• Inventors: Schwarzkopf, Kevin Robert, Gindilis, Andrei L.
• Primary Examiner(s): Menon, Krishnan S
• Assistant Examiner(s): Gerido, Dwan A
• Application Number:
  US12/780,157
• Publication Number:
  US 20110280766A1
• Time in Patent Office: 2,594 Days
• Field of Search: 422 8202, 422 8201, 436501, 436 43, 435 619, 435 61, 435 71
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 27/126 comprising organic polymers
• CPC Class Codes: G01N 27/3276 being a hybridisation with immobilised receptors using a FET type sensor G01N27/4145; concerning the hybridisation C12Q1/68
• CPC Class Codes: G01N 33/0031 comprising two or more sensors, e.g. a sensor array electrochemical electrode arrays G01N27/27
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

Embodiments of the present invention are directed to efficient sensors for detecting the presence and concentration of one or more particular target molecules in solutions, air or other gasses, or otherwise present in an environment or sample. Various embodiments of the present invention provide relatively large binding constants or, equivalently, relatively small dissociation constants for binding of target biopolymers or other target molecules with complementary probes bound to, or associated with, a sensor substrate. In addition, various embodiments of the present invention increase the area of the surface of the sensor affected by binding of a target biopolymer or other target molecules to the sensor which, in turn, produces a stronger signal and greater signal-to-noise ratio for a given number or concentration of bound target biopolymers or other target molecules. In certain embodiments of the present invention, large binding constants, large signal-to-noise ratios, and strong signals are achieved by including probe molecules on the surface of the sensor directed to multiple binding sites within a target biopolymer or other target molecules, so that each bound target biopolymer or other target molecule is bound to, or associated with multiple probes.

Claims

1. A sensor comprising:

a substrate;

a signal-generation component coupled to the substrate that produces a sensor signal; and

two or more types of probes associated with, or bound to, the substrate, each type of probe binding to a different binding site of a target, so that, when the sensor is exposed to the target, the target is bound to two or more probes at two or more binding sites to produce a change, in one or more physical characteristics of the substrate, probes, and/or other substrate-associated entities, that is detected by the signal-generation component, which generates a corresponding sensor signal.

2. The sensor of claim 1

wherein the substrate is the surface of an electrode;

wherein the one or more physical characteristics are modeled as a capacitance associated with the substrate, probes, and/or other substrate-associated entities; and

wherein the signal-generation component detects a change in impedance of a sensor circuit that includes the electrode as a circuit component.

3. The sensor of claim 1

wherein the probes are oligonucleotides bound to the substrate;

and

wherein the target is a nucleotide polymer with multiple subsequences, each subsequence complementary to a different oligonucleotide probe.

4. The sensor of claim 3 wherein:

a first oligonucleotide probe with a first sequence and a second oligonucleotide probe with a second sequence are covalently bound to the substrate, the first sequence complementary to a first subsequence of the target and the second sequence complementary to a second subsequence of the target.

5. The sensor of claim 4 wherein the first oligonucleotide probe is linked through a 3′

end to the substrate and the second oligonucleotide probe is linked through a 5′

end to the substrate.

6. The sensor of claim 4 wherein the first oligonucleotide probe is linked through a 3′

end to the substrate and the second oligonucleotide probe is linked through a 3′

end to the substrate.

7. The sensor of claim 4 wherein the first oligonucleotide probe is linked through a 5′

end to the substrate and the second oligonucleotide probe is linked through a 5′

end to the substrate.

8. The sensor of claim 3 wherein:

a first oligonucleotide probe with a first sequence, a second oligonucleotide probe with a second sequence, a third oligonucleotide probe with a third sequence, and a fourth oligonucleotide probe with a fourth sequence are covalently bound to the substrate, the first sequence and second subsequence complementary to a first subsequence and a second subsequence of a first strand of the target, respectively, and the third sequence and fourth sequence complementary to a third subsequence and fourth subsequence of a second strand of the target.

9. The sensor of claim 3 wherein:

a first oligonucleotide probe with a first oligonucleotide-probe subsequence and a second oligonucleotide-probe subsequence are covalently bound to the substrate, the first oligonucleotide-probe subsequence complementary to a first target subsequence of the target and the second oligonucleotide-probe subsequence complementary to a second target subsequence of the target.

10. The sensor of claim 3 wherein:

a first oligonucleotide probe with a first oligonucleotide-probe subsequence and a second oligonucleotide-probe subsequence is covalently bound to the substrate, the first oligonucleotide-probe subsequence complementary to a first target subsequence of a first strand of the target and the second oligonucleotide-probe subsequence complementary to a second target subsequence of the first strand target; and

a second oligonucleotide probe with a third oligonucleotide-probe subsequence and a fourth oligonucleotide-probe subsequence is covalently bound to the substrate, the third oligonucleotide-probe subsequence complementary to a first target subsequence of a second strand of the target and the fourth oligonucleotide-probe subsequence complementary to a second target subsequence of the second strand target.

11. A method for preparing a sensor, the method comprising:

providing a sensor with a substrate and a signal-generation component coupled to the substrate that produces a sensor signal; and

associating two or more types of probes with, or binding two or more types of probes to, the substrate, each type of probe binding to a different binding site of a target, so that, when the sensor is exposed to the target, the target is bound to two or more probes at two or more binding sites to produce a change, in one or more physical characteristics of the substrate, probes, and/or other substrate-associated entities, that is detected by the signal-generation component, which generates a corresponding sensor signal.

12. The method of claim 11

wherein the substrate is the surface of an electrode;

wherein the one or more physical characteristics are modeled as a capacitance associated with the substrate, probes, and/or other substrate-associated entities; and

wherein the signal-generation component detects a change in impedance of a sensor circuit that includes the electrode as a circuit component.

13. The method of claim 11

wherein the probes are oligonucleotides bound to the substrate;

and

wherein the target is a nucleotide polymer with multiple subsequences, each subsequence complementary to a different oligonucleotide probe.

14. The method of claim 13 further including:

covalently binding a first oligonucleotide probe with a first sequence and a second oligonucleotide probe with a second sequence to the substrate, the first sequence complementary to a first subsequence of the target and the second sequence complementary to a second subsequence of the target.

15. The method of claim 14 further including linking the first oligonucleotide probe through a 3′

end to the substrate and linking the second oligonucleotide probe through a 5′

end to the substrate.

16. The method of claim 14 further including linking the first oligonucleotide probe a 3′

end to the substrate and linking the second oligonucleotide probe through a 3′

end to the substrate.

17. The method of claim 14 further including linking the first oligonucleotide probe through a 5′

end to the substrate and linking the second oligonucleotide probe through a 5′

end to the substrate.

18. The method of claim 14 further including:

covalently binding a first oligonucleotide probe with a first sequence, a second oligonucleotide probe with a second sequence, a third oligonucleotide probe with a third sequence, and a fourth oligonucleotide probe with a fourth sequence to the substrate, the first sequence and second subsequence complementary to a first subsequence and a second subsequence of a first strand of the target, respectively, and the third sequence and fourth sequence complementary to a third subsequence and fourth subsequence of a second strand of the target.

19. The method of claim 13 further including:

covalently binding a first oligonucleotide probe with a first oligonucleotide-probe subsequence and a second oligonucleotide-probe subsequence to the substrate, the first oligonucleotide-probe subsequence complementary to a first target subsequence of the target and the second oligonucleotide-probe subsequence complementary to a second target subsequence of the target.

20. The method of claim 13 further including:

covalently binding a first oligonucleotide probe with a first oligonucleotide probe subsequence and a second oligonucleotide probe subsequence to the substrate, the first oligonucleotide probe subsequence complementary to a first target subsequence of a first strand of the target and the second oligonucleotide probe subsequence complementary to a second target subsequence of the first strand target; and

covalently binding a second oligonucleotide probe with a third oligonucleotide probe subsequence and a fourth oligonucleotide probe subsequence to the substrate, the third oligonucleotide probe subsequence complementary to a first target subsequence of a second strand of the target and the fourth oligonucleotide probe subsequence complementary to a second target subsequence of the second strand target.