Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays
First Claim
1. An assay system, comprising:
- a receptor for receiving a sample solution that may include at least one selected analyte;
a reaction zone comprising at least a portion of said receptor or in fluid communication with said receptor, said reaction zone including at least one reagent configured to produce a reaction mixture with said sample solution;
at least one detection zone in fluid communication with said reaction zone and configured to receive at least a portion of said reaction mixture transported from said reaction zone, said at least one detection zone is carried upon a surface of at least a portion of a waveguide; and
a detector oriented to receive signals from said at least one detection zone and configured to sense a physically detectable change in said at least one detection zone, said physically detectable change correlating with at least one of an absence, a presence, and an amount of said at least one selected analyte in said sample solution, said detector configured to generate a signal correlating with at least one of said absence, said presence, and said amount.
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Abstract
Methods and apparatus for evanescent light fluoroimmunoassays are disclosed. The apparatus employs a planar waveguide with an integral semicylindrical lens, and has multi-analyte features and calibration features, along with improved evanescent field intensity. A preferred embodiment of the biosensor and assay method has patches of capture molecules, each specific for a different analyte disposed adjacently within a single reservoir. The capture molecules are immobilized to the patches on the waveguide surface by site-specific coupling of thiol groups on the capture molecules to photo-affinity crosslinkers, which in turn are coupled to the waveguide surface or to a nonspecific binding-resistant coating on the surface. The patches of different antibodies are produced by selectively irradiating a portion of the waveguide surface during the process of coupling the photo-affinity crosslinkers, the selective irradiation involving a mask, a laser light source, or the like.
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Citations
30 Claims
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1. An assay system, comprising:
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a receptor for receiving a sample solution that may include at least one selected analyte;
a reaction zone comprising at least a portion of said receptor or in fluid communication with said receptor, said reaction zone including at least one reagent configured to produce a reaction mixture with said sample solution;
at least one detection zone in fluid communication with said reaction zone and configured to receive at least a portion of said reaction mixture transported from said reaction zone, said at least one detection zone is carried upon a surface of at least a portion of a waveguide; and
a detector oriented to receive signals from said at least one detection zone and configured to sense a physically detectable change in said at least one detection zone, said physically detectable change correlating with at least one of an absence, a presence, and an amount of said at least one selected analyte in said sample solution, said detector configured to generate a signal correlating with at least one of said absence, said presence, and said amount. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. An assay system, comprising:
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a receptor for receiving a sample solution that may include at least one selected analyte;
a reaction zone comprising at least a portion of said receptor or in fluid communication with said receptor, said reaction zone including at least one reagent configured to produce a reaction mixture with said sample solution, said reaction mixture comprising a reaction product including said at least one selected analyte, said at least one reagent bound thereto, and a physically detectable label that emits light when excited by an evanescent field;
at least one detection zone in fluid communication with said reaction zone and configured to receive at least a portion of said reaction mixture transported from said reaction zone;
a source configured and oriented to cause the generation of said evanescent field at said at least one detection zone; and
a detector oriented to receive signals from said at least one detection zone and configured to sense a physically detectable change in said at least one detection zone, said physically detectable change correlating with at least one of an absence, a presence, and an amount of said at least one selected analyte in said sample solution, said detector configured to generate a signal correlating with at least one of said absence, said presence, and said amount. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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Specification