Method and device for detecting the presence of analyte in a sample

• US 5,451,504 A
• Filed: 07/29/1991
• Issued: 09/19/1995
• Est. Priority Date: 07/29/1991
• Status: Expired due to Term

• Current Assignee: Serex Incorporated (Nymox Pharmaceutical Corporation)
• Original Assignee: Serex Incorporated (Nymox Pharmaceutical Corporation)
• Inventors: Fitzpatrick, Judith, Lenda, Regina B.
• Primary Examiner(s): Scheiner, Toni R.
• Application Number:
  US07/737,091
• Time in Patent Office: 1,513 Days
• Field of Search: 435/7.2, 435/7.9, 435/7.92-7.95, 435/805, 435/968-971, 435/973, 422/55-60, 436/518, 436/523, 436/527, 436/530, 436/531, 436/538, 436/804, 436/810, 436/822
• US Class Current: 435/7.2
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte

Abstract

An assay provides for detecting the presence of an analyte in a sample. Sample is applied to move through three zones. A mobilizeable receptor capable of binding to the analyte is present in the first zone. A trap for unbound receptor, consisting of immobilized ligand, is present in the second zone. Receptor bound to analyte will not bind to the ligand; unbound receptor will bind to the ligand. Mobilization and migration of the receptor can be detected in the third zone, which positively indicates that analyte is present. A device for carrying out the method is also provided.

Claims

1. A method for detecting at least one analyte in a liquid sample comprising:

(a) moving the sample along a pathway comprising three zones located on a single solid phase support, wherein the sample moves from a first zone comprising mobilizeable labeled receptor, which are mobilized by and move with the sample, which receptors specifically bind to and form labeled receptor-analyte complexes with analyte in the presence of analyte in the sample, to a second zone comprising immobilized ligand which binds the mobilized labeled receptors that are not complexed to analye and has a lower association constant than the analyte for the labeled receptor so that the immobilized ligand does not affect the stability of any labeled receptor-analyte complex formed by the moving sample and the labeled receptor and into a third zone for detecting the presence of the labeled receptor-analyte complex; and

(b) detecting the presence of labeled receptor-analyte complexes in the third zone, which positively indicates the presence of analyte in the sample.

3. The method of claim 1 or 2 in which the solid phase support is selected from the group consisting of coated plastic, coated glass, polymer beads, a filter, a silica gel, paper and a membrane.

4. The method of claim 1 or 2 in which the receptor is selected from the group consisting of an antibody having one or more binding sites for the analyte and the ligand, a fragment of an antibody having one or more binding sites for the analyte and the ligand, a cell surface receptor having one or more binding sites for the analyte and the ligand, and a fragment of a cell surface receptor having one or more binding sites for the analyte and the ligand.

5. The method of claim 1 or 2 in which the ligand is selected from the group consisting of analyte analogue, derivative, modification and isomer.

6. The method of claim 1 in which the third zone comprises an immobilized binding partner of the receptor.

8. The method of claim 1 or 2 in which the label is selected from the group consisting of an enzyme, a fluorophore, a chromophore, a dye, colloidal gold, a detectable latex particle and a chemiluminescent agent.

9. The method of claim 1 for detecting more than one analyte in the sample in which the first zone comprises more than one receptor, in which a receptor specific for each analyte is present, the second zone comprises more than one ligand, in which a ligand is present for each analyte is present, the second zone comprises more than one receptor, which a receptor is present for each analyte, and the detection zone comprises one binding partner specific for each receptor, wherein each binding partner is on a discrete area of the detection zone.

11. The method of claim 1 or 2 in which the movement of the sample is facilitated by an absorbent zone which comprises an extension of a solid phase support or a hydrogel.

2. A method for detecting at least one analyte in a liquid sample comprising:

(a) moving the sample along a pathway comprising three zones located on a single solid phase support, wherein the sample moves from a first zone comprising mobilizeable labeled ligands which are mobilized by and move with the sample, and have a lower association constant than the analyte for a receptor so that the labeled ligands do not affect the stability of any receptor-analyte complexes formed by the moving sample and receptors, to a second zone comprising receptors immobilized therein, which in the absence of analyte bind to mobilized labeled ligands, but in the presence of analyte bind to the analyte so that mobilized labeled ligands do not bind to the receptors and move into a third zone for detecting the presence of labeled ligands; and

(b) detecting the presence of labeled ligands in the third zone, which positively indicates the presence of analyte in the sample.

7. The method of claim 2 in which the third zone comprises an immobilized binding partner of the labeled ligand.

10. The method of claim 2 for detecting more than one analyte in the sample in which the first zone comprises more than one ligand, in which a ligand for each analyte is present, the second zone comprises more than one receptor, in which a receptor is present for each analyte, and the detection zone comprises one binding partner specific for each ligand, wherein each binding partner is on a discrete area of the detection zone.