Detection of Promiscuous Small Submicrometer Aggregates
First Claim
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1. A method of detecting aggregate-forming particles or promiscuous inhibitor molecules comprising:
- (a) applying test species to a colorimetric resonant reflectance biosensor or a grating-based waveguide biosensor; and
(b) illuminating the biosensor with light and determining peak wavelength value shifts or refractive index changes over time;
wherein, discontinuous, non-linear, or slope of greater than 2 pm/minute peak wavelength value shifts or refractive index changes over time indicates that the test species are aggregate-forming particles or promiscuous binding molecules.
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Abstract
The invention provides methods for the detection of aggregating molecules that are capable of promiscuous or non-specific binding to proteins in a time efficient manner without the use of labels.
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Citations
30 Claims
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1. A method of detecting aggregate-forming particles or promiscuous inhibitor molecules comprising:
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(a) applying test species to a colorimetric resonant reflectance biosensor or a grating-based waveguide biosensor; and (b) illuminating the biosensor with light and determining peak wavelength value shifts or refractive index changes over time; wherein, discontinuous, non-linear, or slope of greater than 2 pm/minute peak wavelength value shifts or refractive index changes over time indicates that the test species are aggregate-forming particles or promiscuous binding molecules. - View Dependent Claims (2, 4, 5, 6, 7, 8)
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3. The method of claim 3, wherein the stoichiometry of a binding reaction between the test species and the one or more specific binding substances, binding partners or linkers greater than about 1:
- 1, thereby indicating that the test species are aggregate-forming particles or promiscuous binding molecules.
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9. A method of detecting non-specific binding of a test species comprising:
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(a) applying test species to a colorimetric resonant reflectance biosensor or a grating-based waveguide biosensor, wherein the biosensor has one or more specific binding substances, binding partners or linkers immobilized on a surface of the biosensor; and (b) illuminating the biosensor with light and determining peak wavelength value shifts or refractive index changes over time; wherein, discontinuous, non-linear, or slope of greater than 2 pm/minute peak wavelength value shifts or refractive index changes over time indicates that the test species is non-specifically binding. - View Dependent Claims (10, 11, 12, 13, 14)
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15. A method of detecting non-specific binding of a test species comprising:
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(a) applying a test species at varying concentrations to two or more locations on a calorimetric resonant biosensor or a grating-based waveguide biosensor, wherein the biosensor has one or more specific binding substances, binding partners, or linkers immobilized to the biosensor surface; and (b) illuminating the biosensor with light and detecting peak wavelength values or refractive index values for each of the two or more locations; wherein, a discontinuous, non-linear, or slope of greater than 2 pm/minute refractive index change with regard to increasing concentration of the test species indicates that the test species is non-specifically binding. - View Dependent Claims (16, 17, 18, 19, 20)
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21. A method of detecting aggregate-forming particles or promiscuous inhibitor molecules comprising:
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(a) applying a test species at varying concentrations to two or more locations on a calorimetric resonant biosensor or a grating-based waveguide biosensor; and (b) illuminating the biosensor with light and detecting peak wavelength values or refractive index values for each of the two or more locations; wherein, discontinuous, non-linear or slope of greater than 2 pm/minute peak wavelength shift or refractive index change with regard to increasing concentration of the test species indicates that the test species is an aggregate-forming particle or a promiscuous inhibitor. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28)
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29. A method of detecting promiscuous inhibitor molecules or aggregate-forming particles comprising:
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(a) applying a ligand to a first location of a biosensor, wherein the biosensor has a target molecule comprising a specific binding site for the ligand immobilized on the first location and a second location of the biosensor; (b) applying a test species to the first and second locations of the biosensor; (c) applying a molecule known to bind to the target molecule at the specific binding site of the target molecule to the first and the second locations of the biosensor; (d) illuminating the biosensor with light and determining peak wavelength value shifts or refractive index changes at steps (a)-(c) and determining stoichiometric ratios of binding reactions at the first and second locations of the biosensor;
thereby detecting promiscuous inhibitor molecules or aggregate-forming particles. - View Dependent Claims (30)
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Specification