Methods for detecting and quantifying analytes using ionic species diffusion

• US 10,386,365 B2
• Filed: 12/07/2016
• Issued: 08/20/2019
• Est. Priority Date: 12/07/2015
• Status: Active Grant

• Current Assignee: Nanohmics, Inc.
• Original Assignee: Nanohmics, Inc.
• Inventors: Savoy, Steve M., Hoover, Kyle W., Mann, Chris W., Mitchell, Daniel R., John, Jeremy J., Greis, Alexander P.
• Primary Examiner(s): Brown, Melanie
• Application Number:
  US15/372,343
• Publication Number:
  US 20170160227A1
• Time in Patent Office: 986 Days
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

Methods and sensors for detection and quantification of one or more analyte in a test sample are described. A response profile of an ion sensor to a control sample of a known interrogator ion is determined. The ion sensor is exposed to a test sample then to a second sample comprising the known interrogator ion, and a test sample response profile of the ion sensor is determined. One or more test sample sensor response profiles are compared with one or more control sensor response profiles for detecting, identifying, and quantifying one or more analytes in the test sample.

Claims

1. A method for determining the presence of an analyte in a test sample, the method comprising the steps of:

(a) exposing an ion sensor, derivatized with analyte binders capable of interacting specifically with the analyte, to a first liquid ionic solution sample comprising a known interrogator ion species and lacking the analyte;

(b) stopping sensor exposure to the first liquid ionic solution sample;

(c) determining a first ion sensor response profile from a selected time prior to initiating sensor exposure to the first liquid ionic solution sample to a selected time after stopping sensor exposure to the first liquid ionic solution sample and during diffusion of the known interrogator ion species to and from the sensor and through the analyte binders;

(d) exposing the sensor to the test sample, under conditions effective for formation of binder-analyte complexes;

(e) terminating exposure of the sensor to the test sample;

(f) after terminating exposure of the sensor to the test sample,

i) exposing the sensor to a second liquid ionic solution sample, the second liquid ionic solution sample comprising the known interrogator ion species,

ii) stopping sensor exposure to the second liquid ionic solution sample, and

iii) determining a second ion sensor response profile from a selected time prior to initiating sensor exposure to the second liquid ionic solution sample to a selected time after stopping sensor exposure to the second liquid ionic solution sample and during diffusion of the known interrogator ion species to and from the sensor; and

(g) indicating a presence of the analyte in the test sample if the first and second ion sensor response profiles indicate that diffusion of the known interrogator ion species to and from the sensor and through the analyte binders is different from diffusion of the known interrogator ion species to and from the sensor after terminating exposure of the ion sensor to the test sample.

2. The method of claim 1 wherein the first and second liquid ionic solution samples each comprise a plurality of known interrogator ion species and wherein the known interrogator ion species are the same in the first and second liquid ionic solution samples.

3. The method of claim 1 further comprising comparing the first and second ion sensor response profiles by performing a ratiometric comparison of the profiles.

4. The method of claim 1 wherein the analyte binders are biomolecules.

5. The method of claim 1 wherein the analyte binders are ionophores.

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

7. The method of claim 6 wherein the test sample comprises a biological sample.

8. The method of claim 6, wherein the biomolecule is a biomarker.

9. The method of claim 8, wherein the biomarker is an ion.

10. The method of claim 1 wherein the test sample comprises a liquid.

11. The method of claim 1 wherein the ion sensor is in a sensor array.

12. The method of claim 1 wherein the ion sensor is a conductometric semiconducting metal oxide sensor.

13. The method of claim 12 wherein the ion sensor comprises a plurality of nanotraces made by nanoimprint lithography.

14. The method of claim 13 wherein two or more nanotraces have different widths.

15. The method of claim 1 wherein the analyte binders are attached to the ion sensor with linkers.

16. The method of claim 1 wherein the analyte binders are coupled to a porous matrix.

17. The method of claim 1 further comprising quantifying the analyte.

18. A method for detecting the presence of a plurality of different selected analytes in a test sample, the method comprising:

(a) exposing at least a first ion sensor and a second ion sensor to a first liquid ionic solution sample comprising a known interrogator ion species and lacking at least a first analyte and a second analyte, wherein the first ion sensor is derivatized with selected first analyte binders capable of interacting specifically with the first analyte and the second ion sensor is derivatized with selected second analyte binders capable of interacting specifically with the second analyte;

(b) stopping exposure of the first and second ion sensors to the first liquid ionic solution sample;

(c) determining a first ion sensor response profile from a selected time prior to initiating exposure of the first ion sensor to the first liquid ionic solution sample to a selected time after stopping exposure of the first ion sensor to the first liquid ionic solution sample and during diffusion of the known interrogator ion species to and from the first ion sensor and through the selected first analyte binders;

(d) determining a second ion sensor response profile from a selected time prior to initiating exposure of the second ion sensor to the first liquid ionic solution sample to a selected time after stopping exposure of the second ion sensor to the first liquid ionic solution sample and during diffusion of the known interrogator ion species to and from the second ion sensor and through the selected second analyte binders;

(e) exposing the first ion sensor to the test sample, under conditions effective for formation of first binder-analyte complexes comprising the first analyte and the selected first analyte binders and exposing the second ion sensor to the test sample, under conditions effective for formation of second binder-analyte complexes comprising the second analyte and the selected second analyte binders;

(f) terminating exposure of the first and second ion sensors to the test sample;

(g) after terminating exposure of the first and second ion sensors to the test sample,

i) exposing the first and second ion sensors to a second liquid ionic solution sample, the second liquid ionic solution sample comprising the known interrogator ion species,

ii) stopping exposure of the first and second gas sensors to the second liquid ionic solution sample,

iii) determining a third ion sensor response profile from a selected time prior to initiating exposure of the first ion sensor to the second liquid ionic solution sample to a selected time after stopping exposure of the first ion sensor to the second liquid ionic solution sample and during diffusion of the known interrogator ion species to and from the first ion sensor; and

iv) determining a fourth ion sensor response profile from a selected time prior to initiating exposure of the second ion sensor to the second liquid ionic solution sample to a selected time after stopping exposure of the second ion sensor to the second liquid ionic solution sample and during diffusion of the known interrogator ion species to and from the second ion sensor;

(h) indicating a presence of the first analyte in the test sample if the first and third ion sensor response profiles indicate that diffusion of the known interrogator ion species to and from the first ion sensor and through the selected first analyte binders is different from diffusion of the known interrogator ion species to and from the first ion sensor after terminating exposure of the first ion sensor to the test sample; and

(i) indicating a presence of the second analyte in the test sample if the second and fourth ion sensor response profiles indicate that diffusion of the known interrogator ion species to and from the second ion sensor and through the selected second analyte binders is different from diffusion of the known interrogator ion species to and from the second ion sensor after terminating exposure of the second ion sensor to the test sample.

19. The method of claim 18 wherein the first, second, third, and fourth ion sensor response profiles are stored in a database.

20. The method of claim 19 further comprising determining the quantities of the at least first and second analytes in the test sample.

21. The method of claim 19 further comprising comparing the first and third ion sensor response profiles and comparing the second and fourth ion sensor response profiles using read-out integrated circuits and implementing data analysis deconvolution algorithms.