Binding assay employing labelled reagent
First Claim
1. A binding assay process for determining the concentration of one or more analytes in a liquid sampleusing a capture binding agent having binding sites specific for each analyte expected to be present in the sample and a developing binding material capable of binding to bound analyte, to binding sites of the capture binding agent occupied by bound analyte or to binding sites remaining unoccupied by the analyte,the capture binding agent for a given analyte being immobilized at high density on a support in the form of one or more microspots each having an area less than 1 mm2, and whereinlabelled microspheres having a diameter less than 5 μ
- m being used in the assay in relation to the developing binding material, so that the strength of the signal from the label is representative of the fractional occupancy of the binding sites of the capture binding agent, thereby allowing the concentration of the analyte to be determined.
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Abstract
A binding assay process for an analyte, using a capture binding agent with binding sites specific for the analyte and a developing binding material capable of binding with the bound analyte or with the binding sites on the capture binding agent either occupied by the bound analyte or the remaining unoccupied binding sites, employs the capture binding agent in an amount such that only an insignificant fraction of the sample analyte becomes bound to the capture binding agent, which is preferably provided at high surface density on microspots. A label is used in relation to the developing binding material and is provided by microspheres which are less than 5 μm and carry a marker preferably fluorescent dye molecules. To determine the concentration of sample analyte, the signal strength, which represents the fractional occupancy of the binding sites on the capture binding agent by the analyte, is compared with a dose-response curve computed from standard samples. To detect an analyte comprising a single-stranded DNA sequence the analyte presence is detected by the existence of a signal. A kit for the process comprises the capture binding agent immobilised on a solid support, a developing reagent with the developing binding material attached to the microspheres and, for quantitative assays, standards of known amounts of concentrations of the analyte of interest.
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Citations
16 Claims
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1. A binding assay process for determining the concentration of one or more analytes in a liquid sample
using a capture binding agent having binding sites specific for each analyte expected to be present in the sample and a developing binding material capable of binding to bound analyte, to binding sites of the capture binding agent occupied by bound analyte or to binding sites remaining unoccupied by the analyte, the capture binding agent for a given analyte being immobilized at high density on a support in the form of one or more microspots each having an area less than 1 mm2, and wherein labelled microspheres having a diameter less than 5 μ - m being used in the assay in relation to the developing binding material, so that the strength of the signal from the label is representative of the fractional occupancy of the binding sites of the capture binding agent, thereby allowing the concentration of the analyte to be determined.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A binding assay process for detecting the presence of one or more target nucleic acid sequences in a liquid sample
using one or more capture binding agents comprising an oligonucleotide sequence capable of hybridizing to a given target nucleic acid sequence and a developing binding material comprising an oligonucleotide sequence capable of hybridizing to the bound target nucleic acid sequence, the capture binding agent for a given target nucleic acid sequence being immobilized at high density in the form of one or more microspots each having an area less than 1 mm2, and wherein labelled microspheres having a diameter less than 5 μ - m are used in the assay in relation to the developing binding material, so that the signal from the label indicates the presence of the target nucleic acid sequence, thereby allowing the presence of said one or more target nucleic acid sequences to be detected.
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15. A kit for determining the concentration of one or more analytes in a liquid sample, the kit comprising:
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one or more capture binding agents, each capture binding agent having binding sites specific for a given analyte expected to be present in the sample, wherein the capture binding agents are immobilized at high density on a support in the form of one or more microspots, each microspot having an area less than 1 mm2 ; and
,one or more developing binding materials, each developing binding material being capable of binding to a given bound analyte, to binding sites of a given capture binding agent occupied by bound analyte or to binding sites of a given capture binding agent remaining unoccupied by the analyte; wherein labelled microspheres having a diameter less than 5 μ
m are used in the assay in relation to the developing binding material, so that the strength of the signal from the label is representative of the fractional occupancy of the binding sites of a given capture binding agent, thereby allowing the concentration of the analyte to be determined. - View Dependent Claims (16)
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Specification