Label-free methods for performing assays using a colorimetric resonant reflectance optical biosensor

• US 7,300,803 B2
• Filed: 08/22/2006
• Issued: 11/27/2007
• Est. Priority Date: 10/30/2000
• Status: Expired due to Term

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Sru Biosystems Incorporated
• Inventors: Lin, Bo, Li, Peter, Cunningham, Brian T., Pien, Homer, Pepper, Jane, Qiu, Jean, Gerstenmaier, John
• Primary Examiner(s): Chin; Christopher L.
• Application Number:
  US11/508,597
• Publication Number:
  US 20070054339A1
• Time in Patent Office: 462 Days
• Field of Search: 422/82.05, 422/82.11, 435/5, 435/6, 435/7.2, 435/7.32, 435/7.4, 435/288.3, 435/288.4, 435/288.5, 435/288.7, 436/164, 436/518, 436/524, 436/525, 436/527, 436/805, 436/809, 436/810
• US Class Current: 436/518
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00454 by chemical cleavage from the solid support
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00545 Colours
• CPC Class Codes: B01J 2219/00662 Two-dimensional arrays within two-dimensional arrays
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/0074 Biological products
• 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 21/78 producing a change of colour
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

Methods 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 relates to optical devices.

Claims

1. A method of detecting cleavage of one or more entire specific binding substances from a surface of a colorimetric resonant reflectance optical biosensor, wherein one or more specific binding substances are immobilized on the surface of the biosensor at distinct locations, comprising:

a. detecting a colorimetric resonant reflectance optical biosensor peak wavelength value (PWV) of the distinct locations;

b. applying one or more cleaving molecules to the distinct locations;

c. detecting a colorimetric resonant reflectance optical PWV of the distinct locations; and

d. comparing the PWVs from step (a) with the PWVs from step (c);

wherein the cleavage of one or more entire specific binding substances is detected.

2. The method of claim 1, wherein the colorimetric resonant reflectance optical biosensor comprises an internal surface of a vessel selected from the group consisting of a microtiter well, microtiter plate, test tube, petri dish and microfluidic channel.

3. The method of claim 1, wherein the one or more specific binding substances are immobilized onto the surface of the biosensor via a nickel group, amine group, aldehyde group, acid group, alkane group, alkene group, alkyne group, aromatic group, alcohol group, ether group, ketone group, ester group, amide group, amino acid group, nitro group, nitrile group, carbohydrate group, thiol group, organic phosphate group, lipid group, phospholipid group or steroid group.

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

5. The method of claim 1, wherein the specific binding substance is immobilized on the surface of the colorimetric resonant reflectance optical biosensor by a method selected from the group consisting of physical adsorption, chemical binding, electrochemical binding, electrostatic binding, hydrophobic binding and hydrophilic binding.

6. The method of claim 4, wherein the distinct locations define an array spot of about 50-500 microns in diameter.

7. The method of claim 1, wherein the detection comprises:

a. immobilizing one or more specific binding substances in one or more distinct locations defining an array within a well of a microtiter plate, wherein the distinct locations defining the array are located upon the surface of a colorimetric resonant reflectance optical biosensor which comprises an internal surface of the well;

b. detecting a calorimetric resonant reflectance optical PWV for one or more distinct locations within the well;

c. applying one or more cleaving molecules to the well;

d. detecting a colorimetric resonant reflectance optical PWV for one or more distinct locations within the well; and

e. comparing the PWV from step (b) with the PWV from step (d);

wherein the cleavage of one or more entire specific binding substances at the one or more distinct locations within the well is detected.

8. The method of claim 1, wherein the specific binding substance is selected from the group consisting of nucleic acids, peptides, protein solutions, peptide solutions, single or double stranded DNA solutions, RNA solutions, RNA-DNA hybrid solutions, solutions containing compounds from a combinatorial chemical library, antigen, polyclonal antibody, monoclonal antibody, single chain antibody (scFv), F(ab) fragment, F(ab′

)₂ fragment, Fv fragment, small organic molecule, cell, virus, bacteria, polymer and biological sample.

9. The method of claim 1, wherein a peak wavelength value (PWV) is a relative measure of the binding substance that is bound to the biosensor.

10. The method of claim 1, wherein the cleaving molecule is selected from the group consisting of proteases, lipases, nucleases, lyases, peptidases, hydrolases, ligases, kinases and phosphatases.