METHODS FOR DETECTING TARGET ANALYTES AND ENZYMATIC REACTIONS
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Abstract
A microsphere-based analytic chemistry system and method for making the same is disclosed in which microspheres or particles carrying bioactive agents may be combined randomly or in ordered fashion and dispersed on a substrate to form an array while maintaining the ability to identify the location of bioactive agents and particles within the array using an optically interrogatable, optical signature encoding scheme. A wide variety of modified substrates may be employed which provide either discrete or non-discrete sites for accommodating the microspheres in either random or patterned distributions. The substrates may be constructed from a variety of materials to form either two-dimensional or three-dimensional configurations. In a preferred embodiment, a modified fiber optic bundle or array is employed as a substrate to produce a high density array. The disclosed system and method have utility for detecting target analytes and screening large libraries of bioactive agents.
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Citations
46 Claims
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1-29. -29. (canceled)
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30. An assay method comprising:
- (i) providing a planar array of particles randomly-positioned with respect to particle type, where different types of particles are differently encoded by incorporation of an optically detectable characteristic into the particles that permits said differently encoded particles to be distinguished, and wherein said differently encoded particles have different binding molecules attached thereto;
(ii) determining the spatial coordinates of particular particle types within the planar array;
(iii) contacting said array of particles with a solution such that, if a particular type of analyte is present in said solution, said analyte binds to the corresponding binding molecules of said particles and the binding of said analyte is indicated by an optically detectable signal;
(iv) determining the presence of any such bound analyte by determining the presence of said optically detectable signal; and
(v) identifying said optically detectable characteristic of particles having binding molecules bound to an analyte, without moving or separating said particles being interrogated from other particles in the array, by correlating the spatial coordinates of said particles with the location of the optically detectable signals in order to identify said binding molecules bound to an analyte. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 45, 46)
- (i) providing a planar array of particles randomly-positioned with respect to particle type, where different types of particles are differently encoded by incorporation of an optically detectable characteristic into the particles that permits said differently encoded particles to be distinguished, and wherein said differently encoded particles have different binding molecules attached thereto;
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43. A method of performing an assay using at least one planar array of particles, the method comprising:
- forming the planar array with a number of differently encoded particles, on a defined area of a substrate but wherein differently-encoded particles are randomly-positioned within said area, said differently encoded particles having different analyte binding molecules attached thereto, said particle types being encoded by incorporation of an optically detectable characteristic into the particles, that distinguishes said differently encoded particles and the binding molecules attached thereto;
determining the spatial coordinates of particular particle types within the planar array;
contacting said plurality of particles with a solution such that, if particular analytes are present in said solution, said analytes bind to the corresponding binding molecules attached to said particles to form a paired entity and said analytes carry an optically detectable characteristic;
detecting said paired entities; and
identifying, the optically detectable characteristic of the particles to which binding molecules are attached which form paired entities by correlating the spatial coordinates of said particles with the spatial coordinates of the optically detectable characteristic in order to identify said binding molecules forming said paired entities. - View Dependent Claims (44)
- forming the planar array with a number of differently encoded particles, on a defined area of a substrate but wherein differently-encoded particles are randomly-positioned within said area, said differently encoded particles having different analyte binding molecules attached thereto, said particle types being encoded by incorporation of an optically detectable characteristic into the particles, that distinguishes said differently encoded particles and the binding molecules attached thereto;
Specification