Flow immunosensor apparatus
First Claim
1. A flow immunosensor for performing a real-time analysis of a sample which is to be analyzed and which is not yet present, said flow immunosensor consisting essentially of;
- an exchanger including;
a chamber having an inlet port for sample introduction and an outlet port, a support medium, comprising a bed having a volume of 0.1 to 0.5 ml, in the chamber, an antibody immobilized on the medium wherein the antibody recognizes with specificity and sensitivity a target, a labelled antigen complexed to said immobilized antibody and capable of being displaced by the target, said labelled antigen saturating the antigen-binding sites of said antibody;
a detection apparatus connected to the exchanger via the outlet port; and
a flow means adapted for flowing liquid through the bed at a rate of 0.1 to 2.0 ml/min, thereby providing a target molecule in said sample insufficient time in the vicinity of said antigen-binding site and said labelled antigen for equilibration.
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Abstract
Target moiety is detected by (a) providing an antibody specific to the target, (b) saturating the binding sites of the antibody with a labelled form of the target, (c) flowing a liquid containing the target past the saturated antibody, thereby (d) allowing the target to displace the labelled antigen, and (e) detecting the displaced labelled antigen with a detector for the label. This assay is performed by a flow immunosensor comprising: a liquid flowing through the immunosensor for moving a sample through the immunosensor; a sample receiver for introducing the sample to the liquid stream; a flow controller for moving the liquid stream through the immunosensor; an exchanger connected to the sample receiving area in which the sample is brought in contact with labelled antigens of the target and where any target present is allowed to displace the labelled antigen; a detection apparatus connected to the exchanger for detecting any labelled antigen which is released; and a disposal means connected to the detection apparatus for disposing of or collecting of waste coming from the detection apparatus.
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Citations
17 Claims
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1. A flow immunosensor for performing a real-time analysis of a sample which is to be analyzed and which is not yet present, said flow immunosensor consisting essentially of;
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an exchanger including;
a chamber having an inlet port for sample introduction and an outlet port, a support medium, comprising a bed having a volume of 0.1 to 0.5 ml, in the chamber, an antibody immobilized on the medium wherein the antibody recognizes with specificity and sensitivity a target, a labelled antigen complexed to said immobilized antibody and capable of being displaced by the target, said labelled antigen saturating the antigen-binding sites of said antibody;
a detection apparatus connected to the exchanger via the outlet port; and
a flow means adapted for flowing liquid through the bed at a rate of 0.1 to 2.0 ml/min, thereby providing a target molecule in said sample insufficient time in the vicinity of said antigen-binding site and said labelled antigen for equilibration. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
a radiation source for exciting the fluorophore to generate light and, a reading means for detecting the quantity of light generated and displaying an indication of the amount of light read. -
9. A sensor according to claim 8 wherein the support medium is selected from the group consisting of activated polysaccharide beads, silica beads, glass beads, hollow fibers and activated polymers.
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10. The flow immunosensor of claim 1, wherein said flow means is adapted for flowing liquid through said bed at a rate of 0.3 to 2.0 ml/min.
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11. The flow immunosensor of claim 10, wherein said bed has volume of 0.1 to 0.2 ml.
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12. A sensor according to claim 1, further including a control exchanger, said control exchanger comprising:
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a second support medium, an antibody, immobilized on the medium, which recognizes a target antigen believed to be absent from said sample, a labelled antigen complexed to said immobilized antibody.
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13. A sensor according to claim 12 wherein each exchanger has a volume of about 0.1 to 0.5 ml.
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14. A sensor according to claim 1 wherein the labels of the labelled antigen are selected from the group consisting of radiolabels and fluorophores.
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15. A sensor according to claim 14 wherein the detector comprises a radioactivity measuring device and a means for displaying the amount of radioactivity detected.
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16. A sensor according to claim 15 further including a control exchanger comprising:
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a second support medium, an antibody, immobilized on the medium, which recognizes a target antigen believed to be absent from said sample. a labelled antigen complexed to said immobilized antibody.
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17. The flow immunosensor of claim 6, wherein said bed has volume of 0.1 to 0.2 ml.
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Specification
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- Resources
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Current Assigneethe united states of america as represented by the secretary of the navy
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Original Assigneethe united states of america as represented by the secretary of the navy
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InventorsWemhoff, Gregory A., Ligler, Frances S., Kusterbeck, Anne W., Gaber, Bruce P.
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Primary Examiner(s)ALEXANDER, LYLE
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Application NumberUS07/860,965Time in Patent Office3,360 DaysField of Search435/7.36, 435/7.1, 435/7.92, 435/7.93, 435/805, 435/968, 435/961, 435/967, 436/518, 436/529, 436/538, 436/541, 436/800, 436/801, 436/804, 422/57, 422/70, 422/82, 422/82.01, 422/82.09US Class Current422/402CPC Class CodesG01N 33/53 Immunoassay; Biospecific bi...G01N 33/54306 Solid-phase reaction mechan...G01N 33/54366 Apparatus specially adapted...Y10S 435/80 Elimination or reduction of...Y10S 435/804 Single cell proteinY10S 435/961 including a step of forming...Y10S 435/968 High energy substrates, e.g...
