Amine activated colorimetric resonant biosensor
First Claim
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1. A biosensor comprising:
- (a) a one dimensional or a two dimensional grating layer comprised of a material having a high refractive index, the grating layer having a surface comprising an amine functionality;
(b) a low refractive index material layer that supports the grating layer, wherein when the biosensor is illuminated at least one resonance grating effect is produced on the reflected radiation spectrum, and wherein the depth and period of the grating are less than the wavelength of the resonant grating effect.
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Abstract
Amine functionalized colorimetric resonant biosensor for binding proteins, peptides, DNAs, cells, small molecules, and other chemical or biological molecules that are of interests in the areas of proteomic, genomic, pharmaceutical, drug discovery, and diagnostic studies. The invention relates to a coating process that provides a high density of active amine binding sites on the grating surface of the colorimetric resonant biosensor. The method uses chemical reagents that do not alter or degrade a plastic biosensor structure. The invention further provides for test methods that verify the presence of amine moieties on the activated surface on the colorimetric resonant biosensor.
287 Citations
47 Claims
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1. A biosensor comprising:
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(a) a one dimensional or a two dimensional grating layer comprised of a material having a high refractive index, the grating layer having a surface comprising an amine functionality;
(b) a low refractive index material layer that supports the grating layer, wherein when the biosensor is illuminated at least one resonance grating effect is produced on the reflected radiation spectrum, and wherein the depth and period of the grating are less than the wavelength of the resonant grating effect. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A biosensor comprising:
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(a) a one-dimensional grating surface comprised of a material having a low refractive index;
(b) a substrate layer that supports the grating surface;
(c) a high refractive index material layer that is applied on top of the low refractive index grating surface wherein the high refractive index material has an amine-functionalized surface opposite of the grating surface; and
(d) one or more specific binding substances immobilized on the surface of the high refractive index layer;
wherein, when the biosensor is illuminated a resonant grating effect is produced on a reflected radiation spectrum, and wherein cross-section of the one-dimensional grating is less than the wavelength of the resonant grating effect. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24. A method for preparing an amine-activated colorometric resonant biosensor comprising:
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(a) preparing a grating layer comprising a substrate supporting a grating having a surface comprised of a high refractive index material;
(b) exposing the grating layer to an alcoholic silane solution;
(c) rinsing the grating layer with an alcohol; and
(d) drying the calorimetric resonant biosensor. - View Dependent Claims (25, 26, 27, 28, 29, 30)
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- 31. A fluorescence test method for determining the presence of amine groups on the surface of an amine-functionalized calorimetric resonant biosensor, comprising incubating the biosensor with a fluorescent dye solution to form a dyed biosensor and scanning the biosensor with a fluorescence spectrophotometer.
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34. A colorimetric method for determining the presence of amine groups on the surface of an amine-functionalized calorimetric resonant biosensor, comprising
(a) exposing an amine functionalized biosensor and a non-functionalized biosensor to a test solution, (b) incubating the biosensors in a color developing solution; - and
(c) comparing the color developing solution of the amine-functionalized biosensor to the color developing solution of the non-functionalized biosensor to determine the presence of amine groups on the surface of the amine functionalized biosensor. - View Dependent Claims (35, 36, 37, 38, 39)
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40. An indirect binding test method for determining the presence of amine groups on the surface of an amine-functionalized calorimetric resonant biosensor, comprising:
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(a) attaching a biomolecule to the amine functionalized biomolecule;
(b) incubating the biosensor with a fluorescently labeled binding partner for the biomolecule; and
(c) scanning the biosensor to determine a fluorescence reading, thereby determining the presence of amine groups on the biosensor. - View Dependent Claims (41, 42, 43, 44, 45, 46, 47)
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Specification