Differential separation assay
First Claim
1. A diagnostic system for determining presence of an analyte comprising,an excess amount of fluorescence labeled binding agent forming a complex with a specific target analyte to be determined, the combination having free fluorescence labeled binding agent and bound analyte,an elongated migration path for the bound analyte and the labeled binding agent, the bound analyte and the labeled binding agent having different migration rates, said path having a migration start zone and a downstream migration analysis zone,means for effecting the migration of the bound analyte and the free labeled binding agent,optical means in communication with the analysis zone for timing the migration of fluorescence labeled binding agent and bound analyte from the start zone to the analysis zone, andmeans for comparing the migration times or rates of fluorescence labeled binding agent and bound analyte, whereby the presence of analyte is determined.
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Accused Products
Abstract
An electrophoresis-based assay system for detection of one or more target substances, i.e. an analyte tagged with fluorescent binding agents. The analyte is reacted with an excess amount of fluorescently tagged binding agent. The reaction mixture is subjected to electrophoresis and the migration of bound and free fluorescent substances are timed at a location where there is a spatial and optical differentiation of the two substances. An optical detector supplies signals corresponding to fluorescent amplitudes of the two substances. The free fluorescent substance arrives at a time expected from calibration runs. This optical signal is a marker for a second time, either earlier or later, when the bound substance should have arrived. Recorded data is searched to establish the relation between free and bound dye among the recorded optical signals. An absence of a bound dye signal infers the absence of target analyte in a sample. The amounts of bound and unbound amounts of the same fluorescent substance may be related by ratio of amplitudes of the optical signals so that the amount of target analyte may be estimated.
46 Citations
13 Claims
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1. A diagnostic system for determining presence of an analyte comprising,
an excess amount of fluorescence labeled binding agent forming a complex with a specific target analyte to be determined, the combination having free fluorescence labeled binding agent and bound analyte, an elongated migration path for the bound analyte and the labeled binding agent, the bound analyte and the labeled binding agent having different migration rates, said path having a migration start zone and a downstream migration analysis zone, means for effecting the migration of the bound analyte and the free labeled binding agent, optical means in communication with the analysis zone for timing the migration of fluorescence labeled binding agent and bound analyte from the start zone to the analysis zone, and means for comparing the migration times or rates of fluorescence labeled binding agent and bound analyte, whereby the presence of analyte is determined.
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11. A diagnostic system for detecting target substances during electrophoresis comprising,
an electrophoresis apparatus having a voltage supply terminal, a gel lane and a sample supply source, the voltage supply terminal establishing a path of migration for electrophoretically mobile substances, first optical means including a focused beam, a beam restricting slit or pinhole disposed in the path of the focused beam, and first optical elements projecting an image of the slit or pinhole on the gel lane in said path between the voltage supply terminal and the sample supply source, a light detector isolated from light of the beam, second optical mans including a reflected beam which is the reflection of the focused beam, a filter and second optical elements projecting a reflected image of the slit onto said detector, said first and second optical elements sharing a focusing lens, a sample of free fluorescent substance and analyte material tagged with fluorescent substance, both being mobile substances in said gel lane, means for measuring and recording times of signals representing detection of fluorescent substances, and a data reader means for correlating the presence of free fluorescent substance with bound fluorescent substance.
Specification