Patent AT167179B successfully refreshed from Google Patents.
Patent EP0250137A2 successfully refreshed from Google Patents.
EP0250137A2
Expired / Lapsed
EP
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Colloidal gold immunoassay
Assignee: Ortho Clinical Diagnostics Inc
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Filing: 1987-06-08
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Granted: —
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Expires: 2007-06-08
Abstract
Abstract A new immunoassay method for providing sensitive and rapid detection of ligands in body fluids utilizing colloidal gold labeled reagents and a matrix membrane strip having an immunologically active component immobilized therein. The sample and the fluid reagent having the colloidal gold labeled biologically active molecule is mixed and then brought into contact with one end of the membrane strip allowing flow upward through the membrane. The height of the visually observable color may be correlated to the presence of the ligand to be determined in the fluid sample.
Inventors
- 👤 Dennis M. Mochnal
- 👤 Henry A. Graham
- 👤 Jemo Kang
- 👤 Chi-Deu Chang
- 👤 Shin Chen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-01 22:01
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/585; G01N33/53; G01N33/543; G01N33/54366; G01N33/54386; G01N33/74
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
134 words
A method for detecting a ligand in a sample comprising the steps of: a) providing a membrane strip having a tortuous path therethrough, said membrane having two ends and further having attached thereto a ligand binding partner; b) providing a liquid reagent containing a ligand binding partner or ligand labelled directly or indirectly with colloidal gold; c) contacting one end of said membrane strip with said sample and, simultaneously or successively, with said liquid reagent whereby an immunological reaction is carried out as the sample and the reagent move from said one end to the other end of the membrane strip; and d) observing said membrane for the presence of color, the distance of which from said one end being correlatable to the presence of colloidal gold and of said ligand in said sample.
Claim 2
Independent Claim
134 words
A method for detecting a ligand in a sample comprising the steps of: a) providing a membrane strip having a tortuous path therethrough, said membrane having two ends and further having attached thereto a ligand binding partner; b) providing a liquid reagent containing a ligand binding partner or ligand labelled directly or indirectly with colloidal gold; c) contacting one end of said membrane strip with said sample and, simultaneously or successively, with said liquid reagent whereby an immunological reaction is carried out as the sample and the reagent move from said one end to the other end of the membrane strip; and d) observing said membrane for the presence of color, the distance of which from said one end being correlatable to the presence of colloidal gold and of said ligand in said sample.
Claim 3
Dependent Claim
54 words
The method of claim 1, wherein: in step b) is a ligand, labelled directly or indirectly with colloidal gold, which competes with the ligand in the sample for the ligand binding partner is used; and the reagent and the sample are mixed prior to their simultaneous application the one end of the strip membrane.
Claim 4
Dependent Claim
80 words
The method of claim 1, wherein the ligand is an antigen having a single epitopic site; the ligand binding partner in step a) is an antigen having the same immunological reactivity as said sample antigen; the ligand binding partner in step b) is an antibody, labelled directly or indirectly with colloidal gold, which is specific for said sample antigen; and the reagent and the sample are mixed prior to their simultaneous application to the one end of the strip membrane.
Claim 5
Dependent Claim
51 words
The method of claim 1, wherein: the ligand is an antibody; the ligand binding partner in step a) is an antigen for which the antibody is specific; and the ligand binding partner in the reagent is an immunoglobulin, labelled directly or indirectly with colloidal gold, which is specific for said antibody.
Claim 6
Dependent Claim
67 words
The method of claim 1, wherein: the ligand is an antigen having at least two epitopic sites; the ligand binding partner in step a) is an antibody which is specific for a first epitopic site on the antigen; and the ligand binding partner in step b) is an antibody, labelled directly or indirectly with colloidal gold, which is specific for a second epitopic site on the antigen.
Claim 7
Dependent Claim
80 words
The method of claim 1, wherein: the ligand is an antigen having at least two epitopic sites; the ligand binding partner in step a) is an antigen having the same immunological reactivity as said sample antigen; the ligand binding partner in step b) is an antibody, labelled directly or indirectly with colloidal gold, which is specific for the antigens; and the reagent and the sample are mixed prior to their simultaneous application to the one end of the strip membrane.
Claim 8
Independent Claim
21 words
The method of any one of claims 1 to 3, 5 and 6 wherein the sample antigen is lutinising hormone (LH).
Claim 9
Independent Claim
41 words
The method of any one of claims 1 to 7, wherein said colloidal gold has an optical density ratio of 1.7 to 3.0 measured as the ratio of the optical density at 540 nm to the optical density at 600 nm.
Claim 10
Dependent Claim
12 words
The method of claim 8, wherein the ratio is 2 to 2.5.
Claim 11
Independent Claim
25 words
The method of any one of claims 1 to 9, wherein the membrane strip comprises activated Nylon 66, activated bibulous paper, or activated polyvinylidine difluoride.
Claim 12
Independent Claim
32 words
The method of any one of claims 1 to 10, wherein the one end of the membrane strip is conctacted with a wash solution after it has been contacted with the sample.
Claim 13
Independent Claim
36 words
The method of any one of claims 1 to 11, further comprising the step of incubating said membrane at room temperature for a period of time less than about 10 minutes prior to said observing step.
Claim 14
Independent Claim
52 words
The method of any one of claims 1 to 12, wherein the entire method is performed in a time period equal to the time for migration through the membrane strip, the time required for the contacting steps, the latter generally totalling less than about 30 seconds, and, if used, the incubation time.
Claim 15
Independent Claim
24 words
The method of any one of claims 1 to 13, wherein said membrane strip further comprises a calibrator for said ligand to be detected.
Claim 16
Independent Claim
35 words
The method of any one of claims 1 to 14, wherein said membrane strip has a plurality of different ligand binding partners attached thereto in separate zones whereby a plurality of ligands may be detected.
Claim 17
Dependent Claim
76 words
A kit for use in an assay for the detection of a ligand in a fluid sample comprising: a membrane having immobilized therethrough continuously or in separate zones a ligand binding partner for binding to the ligand; and a container having a colloidal gold labelled ligand or ligand binding partner for binding to the ligand binding partner on the membrane in a competitive assay or to the ligand in the fluid sample in a sandwich assay.
Claim 18
Dependent Claim
76 words
A kit for use in an assay for the detection of a ligand in a fluid sample comprising: a membrane having immobilized therethrough continuously or in separate zones a ligand binding partner for binding to the ligand; and a container having a colloidal gold labelled ligand or ligand binding partner for binding to the ligand binding partner on the membrane in a competitive assay or to the ligand in the fluid sample in a sandwich assay.
References Cited (Backward Citations)
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