Optical analyte detection systems and methods of use

• US 9,921,165 B2
• Filed: 11/04/2011
• Issued: 03/20/2018
• Est. Priority Date: 11/05/2010
• Status: Active Grant

• Current Assignee: Board of Trustees of The University of Illinois, Genalyte, Inc.
• Original Assignee: Board of Trustees of The University of Illinois, Genalyte, Inc.
• Inventors: Bailey, Ryan C., Qavi, Abraham J., Kindt, Jared T., Byeon, Ji-Yeon, Luchansky, Matthew S., McClellan, Melinda S., Washburn, Adam L., Owen, Tate, Gleeson, Martin Anthony, Gunn, III, Lawrence C.
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US13/883,280
• Publication Number:
  US 20130295688A1
• Time in Patent Office: 2,328 Days
• Field of Search: 356128, 356337, 356481, 385 12, 435 611
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 2021/7789 Cavity or resonator
• CPC Class Codes: G01N 21/75 Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated systems in which material is burnt in a flame or plasma G01N21/72, G01N21/73
• CPC Class Codes: G01N 21/7746 the waveguide coupled to a cavity resonator
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

Various embodiments are drawn to systems and methods for detecting an analyte of interest in a sample including an optical sensor, a capture probe attached to a surface of the optical sensor wherein the capture probe is capable of binding to the analyte to form a duplex or complex, and an antibody capable of binding to the analyte, duplex, or complex. In several embodiments, systems and methods further include a particle attached to the antibody or capable of binding to the antibody. In several embodiments, systems and methods for analyte detection feature one or more of the following: high detection sensitivity and specificity, scalability and multiplex capacity, ability to analyze large analytes, and ability to detect or measure multiple individual binding events in real-time.

Claims

1. A system for detecting a polypeptide of interest in a sample comprising:

an optical sensor that comprises a ring resonator;

a plurality of first antibodies that specifically bind to the polypeptide of interest, wherein the first antibodies are attached to a surface of the ring resonator;

a plurality of second antibodies that specifically bind to the polypeptide of interest;

a plurality of particles attached to the second antibodies or particles capable of binding the second antibodies; and

a detector,

wherein said optical sensor has a resonant wavelength that is shifted when said second antibodies are bound to said polypeptide of interest bound to said first antibodies, such that said optical sensor is configured to sense said second antibodies combined with said polypeptide bound to said first antibodies, and the particles amplify the shift of the resonant wavelength, and

wherein the detector is configured to determine the concentration of the polypeptide in the sample based on the shift of the resonant wavelength resulting from multiple bound particles at the ring resonator.

2. A system for detecting a polypeptide of interest in a sample comprising:

an optical sensor that comprises a ring resonator configured to resonate at a resonant wavelength;

a tunable laser capable of providing light at said resonant wavelength for the optical sensor;

a plurality of first antibodies that specifically bind to the polypeptide of interest, wherein the first antibodies are attached to a surface of the ring resonator;

a plurality of second antibodies that specifically bind to the polypeptide of interest;

a plurality of particles attached to the second antibodies or particles capable of binding the second antibodies; and

a detector,

wherein said optical sensor has a resonant wavelength that is shifted when said second antibodies bind to said polypeptide bound to said first antibodies such that said optical sensor is configured to sense said second antibodies combined with said polypeptide bound to the first antibodies;

wherein the particle amplifies the shift of the resonant wavelength;

wherein the detector is capable of detecting the shifted resonant wavelength, and

wherein the detector is configured to determine the concentration of the polypeptide in the sample based on the shift of the resonant wavelength resulting from multiple bound particles at the ring resonator.

3. A system for detecting an analyte of interest in a sample comprising:

an optical sensor that comprises a ring resonator;

a plurality of capture probes attached to a surface of the ring resonator, wherein the capture probes are capable of binding to the analyte;

a plurality of antibodies capable of specifically binding to the analyte or a plurality of complexes formed between the analyte and the capture probes;

a plurality of particles attached to the antibodies or capable of binding to the antibodies; and

a detector,

wherein said optical sensor has a resonant wavelength that is shifted by said antibodies bound to said complexes or analyte bound to the capture probes, such that said optical sensor is configured to sense said antibodies combined with said analyte or complexes, and the particles amplify the shift of the resonant wavelength, and

wherein the detector is configured to determine the concentration of the analyte in the sample based on the shift of the resonant wavelength resulting from multiple bound particles at the ring resonator.

4. The system of claim 3, wherein the particles comprise a bead, polypeptide, nanoparticle, semiconductor crystal, titanium-oxide crystal, or quantum dot.

5. The system of claim 3, wherein the particles comprise an average diameter of at least 1 nm.

6. The system of claim 4, wherein the bead comprises silicon, polystyrene, agarose, sepharose, metal, or metal-oxide.

7. The system of claim 3, wherein the particles comprise a polypeptide of at least 200 Daltons (Da).

8. The system of claim 3, wherein the analyte is a polypeptide.

9. The system of claim 8, wherein the capture probes comprise an antibody that specifically binds to the polypeptide, an aptamer that specifically binds to the polypeptide, or a protein that specifically binds to the polypeptide.

10. The system of claim 3, wherein the analyte is a nucleic acid.

11. The system of claim 10, wherein the nucleic acid comprises ribonucleic acid (RNA).

12. The system of claim 10, wherein the capture probes comprise a DNA oligonucleotide.

13. The system of claim 3, wherein the optical sensor comprises a silicon waveguide structure.

14. The system of claim 13, wherein the optical sensor has an output portion configured to output an optical signal, and wherein the optical output yields different outputs when said capture probe binds to the analyte of interest forming said complex and said antibody binds said complex, and when said antibody does not bind to said complex.

15. The system of claim 3, wherein the ring resonator comprises an input portion and an output portion, and wherein the optical sensor further comprises an input waveguide and an output waveguide having the ring resonator therebetween configured to increase coupling of said light from said input waveguide to said output waveguide when said capture probes bind to the analyte of interest forming said complexes and said antibodies bind to said complexes.

16. The system of claim 3, wherein said optical sensor is integrated on an integrated optical chip comprising optical waveguides.

17. A method for detecting a polypeptide of interest in a sample comprising:

providing an optical sensor that comprises a ring resonator, the optical sensor comprising a plurality of first antibodies attached to a surface of the ring resonator, wherein the first antibodies specifically bind to the polypeptide of interest;

applying a sample for which the presence or absence of the polypeptide of interest is to be determined to the optical sensor, under conditions in which the polypeptide of interest, when present, and the first antibodies bind;

providing a plurality of second antibodies that specifically bind the polypeptide of interest, wherein binding between the second antibodies and the polypeptide of interest, when bound to the first antibodies, shifts a resonant wavelength of the optical sensor;

providing a plurality of particles attached to the second antibodies or particles capable of binding the second antibodies, wherein the particles amplify the shift of the resonant wavelength; and

determining the concentration of the polypeptide of interest in the sample based on the shift of the resonant wavelength resulting from multiple bound particles at the ring resonator.

18. The method of claim 17, wherein said ring resonator comprises a waveguide structure.

19. A method for detecting an analyte of interest in a sample comprising:

providing an optical sensor that comprises a ring resonator, the optical sensor comprising a plurality of capture probes attached to a surface of the ring resonator, wherein the capture probes are capable of binding to the analyte of interest to form complexes;

applying a sample for which the presence or absence of the analyte of interest is to be determined to the optical sensor, under conditions in which the analyte of interest, when present, and the capture probes bind to form the complexes;

providing plurality of antibodies that specifically bind to the complexes or analyte, wherein binding between the antibodies and the complexes or the analyte, when the analyte is bound to the capture probes, shifts a resonant wavelength of the optical sensor;

providing a plurality of particles attached to the antibodies or particles capable of binding the antibodies, wherein the particles amplify the shift of the resonant wavelength; and

determining the concentration of the analyte of interest in the sample based on the shift of the resonant wavelength resulting from multiple bound particles at the ring resonator.

20. The method of claim 19, wherein the analyte of interest comprises ribonucleic acid (RNA).

21. The method of claim 19, wherein said ring resonator comprises a waveguide structure.