Optical detection of label-free biomolecular interactions using microreplicated plastic

• US 8,580,578 B2
• Filed: 04/22/2011
• Issued: 11/12/2013
• Est. Priority Date: 10/30/2000
• Status: Expired due to Term

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Inventors: Cunningham, Brian T., Pepper, Jane, Lin, Bo, Li, Peter, Pien, Homer, Qiu, Jean
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US13/092,296
• Publication Number:
  US 20110201132A1
• Time in Patent Office: 935 Days
• Field of Search: None
• US Class Current: 436/524
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/4788 Diffraction for sizing particles G01N15/0205
• CPC Class Codes: G01N 21/7743 the reagent-coated grating coupling light in or out of the waveguide
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G02B 5/1809 with pitch less than or comparable to the wavelength
• CPC Class Codes: G02B 5/1852 using mechanical means, e.g. ruling with diamond tool, moulding
• CPC Class Codes: G02B 6/02061 Grating external to the fibre and in contact with the fibre, e.g. evanescently coupled, gratings applied to the fibre end mechanically induced in the fibre G02B6/02071, surface relief on the fibre G02B6/02066
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/813 Cancer

Abstract

Methods and compositions are provided for detecting biomolecular interactions. The use of labels is not required and the methods can be performed in a high-throughput manner. The invention also provides optical devices useful as narrow band filters.

Claims

1. A method of determining the amount of one or more binding partners in a test sample comprising:

(a) illuminating a biosensor with light, wherein the biosensor comprises an optical grating;

a substrate layer that supports the optical grating;

the optical grating being replicated from a master grating structure by causing a liquid to harden after dispensing the liquid between the master grating structure and the substrate layer, wherein the optical grating comprises the hardened liquid and wherein the optical grating comprises features having a depth and a cross-sectional period; and

one or more specific binding substances immobilized on a surface of the optical grating;

wherein, when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum, and wherein the depth and cross-sectional period of the optical grating are less than the wavelength of the resonant grating effect;

(b) applying a test sample comprising one or more binding partners to the biosensor;

(c) illuminating the biosensor with light; and

(d) detecting reflected wavelength of light from the biosensor; and

(e) (i) comparing the reflected wavelength of light from the biosensor to positive and negative controls;

or (ii) comparing the reflected wavelength of light from the biosensor to a reflected wavelength of light detected from the biosensor prior to the applying the test sample to the biosensor, wherein the amount of one or more binding partners in a test sample is detected.

2. The method of claim 1, wherein a narrow band of optical wavelengths is reflected from the biosensor when the biosensor is illuminated with a broad band of optical wavelengths.

3. The method of claim 1, wherein the substrate layer comprises plastic.

4. The method of claim 1, wherein the hardened liquid comprises hardened optical cement.

5. The method of claim 1, wherein the surface of the optical grating opposite the substrate layer is coated with a material having a refractive index higher than the substrate layer, and wherein the one or more specific binding substances are immobilized on the surface of the material having a refractive index higher than the substrate layer opposite the optical grating.

6. The method of claim 5, wherein the material having a refractive index higher than the substrate layer is selected from the group consisting of zinc sulfide, titanium dioxide, tantalum oxide, and silicon nitride.

7. The method of claim 1 further comprising a cover layer on the surface of the optical grating opposite the substrate layer, wherein the one or more specific binding substances are immobilized on the surface of the cover layer opposite the optical grating.

8. The method of claim 7, wherein the cover layer comprises a material that has a lower refractive index than the optical grating.

9. The method of claim 8, wherein the cover layer comprises a material selected from the group consisting of glass, epoxy, and plastic.

10. The method of claim 1, wherein the optical grating comprises a repeating pattern having a cross-sectional period of about 0.01 microns to about 1 micron and a depth of about 0.01 microns to about 1 micron.

11. The method of claim 1, wherein the one or more specific binding substances are arranged in an array of distinct locations.

12. The method of claim 1, wherein the one or more specific binding substances are immobilized on the optical grating by physical adsorption or by chemical binding.

13. The method of claim 1, wherein the one or more specific binding substances are bound to their binding partners.

14. The method of claim 13, wherein the binding partners are selected from the group consisting of proteins, peptides, single strand DNA, double strand DNA, RNA, chemical molecules in solution, nucleic acids, polypeptides, antigens, polyclonal antibodies, monoclonal antibodies, single chain antibodies (scFv), F(ab) fragments, F(ab′

)₂ fragments, Fv fragments, small organic molecules, cells, viruses, bacteria, and biological samples.

15. The method of claim 1, wherein the one or more specific binding substances are selected from the group consisting of protein solutions, peptide solutions, DNA solutions, RNA solutions, solutions of combinatorial chemical libraries, nucleic acids, polypeptides, antigens, polyclonal antibodies, monoclonal antibodies, single chain antibodies (scFv), F(ab) fragments, F(ab′

)₂ fragments, Fv fragments, small organic molecules, cells, viruses, bacteria, and biological samples.

16. The method of claim 1, further comprising an antireflective dielectric coating on the surface of the substrate opposite of the optical grating.

17. The method of claim 1, wherein the biosensor is attached to a bottomless microtiter plate.

18. The method of claim 1, wherein the optical grating is comprised of a repeating pattern of shapes selected from the group consisting of continuous parallel lines, squares, circles, ellipses, triangles, ovals, trapezoids, sinusoidal waves, rectangles, and hexagons.

19. The method of claim 1, wherein the optical grating comprises an optically transparent material that conducts electricity, wherein the surface of the optical grating is coated with an electrical insulator.

20. The method of claim 1, wherein the surface of the optical grating is amine activated, aldehyde activated, or nickel activated and the one or more specific binding substances are immobilized on the surface of the amine activated, aldehyde activated, or nickel activated optical grating.

21. The method of claim 20, wherein the one or more specific binding substances are immobilized on the surface of the amine activated, aldehyde activated, or nickel activated optical grating via a chemical linker.