EP0245926A2
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EP
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Method of measuring antigens or antibodies in biological fluids using ligand labeled antigens or ligand labeled antibodies
Assignee: Siemens Medical Solutions Diagnostics Inc Canada
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Filing: 1987-03-06
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Granted: —
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Expires: 2007-03-06
Abstract
Abstract A method for measuring the level of circulating antigen (Ag₁), antibody (Ab₁) or hapten (H) in a liquid sample, which comprises forming in a liquid phase reaction a soluble complex wherein said antigen (Ag₁) antibody (Ab₁) or hapten (H) is linked through, respectively, a specific antibody (Ab), antigen (Ag) or antihapten (Anti-H), to a matrix which is soluble in the liquid phase and carries a ligand (X), subsequently forming an insolubilised complex comprising a solid support linked to the ligand (X) of said soluble complex through an anti-ligand (Y), the insolubilised complex carrying a label (Z) linked to the antigen (Ag₁) through an anti-antigen (Anti-Ag₁), to the antibody (Ab₁) through an anti-antibody (Anti-Ab₁) or to the hapten (H), washing the insolubilised complex, and checking the washed insolubilised complex for the presence of the label (Z) to provide a measure of the level of said antigen (Ag₁), antibody (Ab₁) or hapten (H) in said sample.
Inventors
- 👤 A. Said El Shami
- 👤 Olusola O. Alaba
- 👤 Charles A. Kasal
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 01:50
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54306; G01N33/531; G01N33/536; G01N33/537; G01N33/538; G01N33/544; G01N33/548; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/971; Y10S436/00; Y10S436/823
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
155 words
A method for measuring the level of an antigen (Ag₁), antibody (Ab₁) or hapten (H) in a liquid sample, which comprises forming in a liquid phase reaction a soluble complex wherein said antigen (Ag₁) antibody (Ab₁) or hapten (H) is linked through, respectively, a specific antibody (Ab), antigen (Ag) or antihapten (Anti-H), to a matrix which is soluble in the liquid phase and carries a ligand (X), subsequently forming an insolubilised complex comprising a solid support linked to the ligand (X) of said soluble complex through an anti-ligand (Y), the insolubilised complex carrying a label (Z) linked to the antigen (Ag₁) through an anti-antigen (Anti-Ag₁), to the antibody (Ab₁) through an anti-antibody (Anti-Ab₁) or to the hapten (H), washing the insolubilised co mplex, and checking the washed insolubilised complex for the presence of the label (Z) to provide a measure of the level of said antigen (Ag₁), antibody (Ab₁) or hapten (H) in said sample.
Claim 2
Independent Claim
155 words
A method for measuring the level of an antigen (Ag₁), antibody (Ab₁) or hapten (H) in a liquid sample, which comprises forming in a liquid phase reaction a soluble complex wherein said antigen (Ag₁) antibody (Ab₁) or hapten (H) is linked through, respectively, a specific antibody (Ab), antigen (Ag) or antihapten (Anti-H), to a matrix which is soluble in the liquid phase and carries a ligand (X), subsequently forming an insolubilised complex comprising a solid support linked to the ligand (X) of said soluble complex through an anti-ligand (Y), the insolubilised complex carrying a label (Z) linked to the antigen (Ag₁) through an anti-antigen (Anti-Ag₁), to the antibody (Ab₁) through an anti-antibody (Anti-Ab₁) or to the hapten (H), washing the insolubilised co mplex, and checking the washed insolubilised complex for the presence of the label (Z) to provide a measure of the level of said antigen (Ag₁), antibody (Ab₁) or hapten (H) in said sample.
Claim 3
Dependent Claim
27 words
A method according to claim 1 wherein the label (Z) is an enzymatic label, a radioactive label, a fluorescent label, a chemiluminescent label or a bioluminescent label.
Claim 4
Dependent Claim
38 words
A method according to claim 1 or 2 wherein the matrix is a soluble carbohydrate with a molecular mass of from 6,000 to 80,000 daltons or a Ficoll with a molecular mass of from 70,000 to 400,000 daltons.
Claim 5
Dependent Claim
12 words
A method according to claim 3 wherein the matrix is a dextran.
Claim 6
Dependent Claim
27 words
A method according to claim 1 or 2 wherein the matrix is a soluble polymer or copolymer with a molecular mass of from 5,000 to 200,000 daltons.
Claim 7
Independent Claim
25 words
A method according to any one of the preceding claims wherein the ligand (X) is covalently attached to the matrix through an amino acid copolymer.
Claim 8
Independent Claim
43 words
A method according to any one of the preceding claims wherein the ligand (X) is avidin, the solid support is a coated tube coated with avidin, and biotin is added to effect the immobilization of the soluble complex onto the avidin coated tube.
Claim 9
Independent Claim
46 words
A method according to any of claims 1 to 6 wherein the ligand (X) is biotin, the solid support is a coated tube coated with an anti-avidin antibody, and avidin is added to effect the immobilization of the soluble complex onto the anti-avidin antibody coated tube.
Claim 10
Independent Claim
45 words
A method according to any one of claims 1 to 6 wherein the ligand is biotin, the solid support is a coated tube coated with biotinylated protein, and avidin is added to effect the immobilization of the soluble complex onto the biotinylated protein coated tube.
Claim 11
Independent Claim
42 words
A method according to any one of the preceding claims wherein the ligand is selected from hapten, biotin, fluoresceine, dinitrophenol, avidin and bovine serum albumin, and the anti-ligand is selected from anti-hapten, avidin, anti-biotin, anti-fluoresceine, anti-dinitrophenol, anti-avidin and anti-bovine serum albumin, respectively.
Claim 12
Independent Claim
34 words
A method according to any one of the preceding claims wherein the ligand (X) is covalently attached to the matrix, and the specific antibody (Ab) or antigen (Ag) is chemically attached to the matrix.
Claim 13
Independent Claim
100 words
A method according to any one of the preceding claims wherein the sample contains an antigen or antibody, the soluble complex is formed by reacting the sample in the liquid phase with a said matrix carrying ligand (X) and specific antibody (Ab) or antigen (Ag) in the presence of an anti-antigen (Anti-Ag₁) or anti-antibody (Anti-Ab₁) labelled with said label (Z) to form a said soluble complex, reacting this soluble complex with immobilized anti-ligand (Y) on a solid support to form an insolubilised complex, washing the insolubilised complex, and checking for the label (Z) on the anti-antigen (Anti-Ag₁) or anti-antibody (Anti-Ab₁).
Claim 14
Independent Claim
112 words
A method according to any one of claims 1 to 11 wherein the sample contains an antigen (Ag₁) or antibody (Ab₁), the soluble complex is formed by reacting the sample in the liquid phase with a said matrix carrying said ligand (X) and said specific antibody (Ab) or antigen (Ag), the insolubilised complex is formed by contacting said soluble complex with immobilized anti-ligand (Y) on a solid support to form an insolubilised complex, washing the insolubilised complex, reacting it with an anti-antigen (Anti-Ag₁) or anti-antibody (Anti-Ab₁) labelled with said label (Z), rewashing the resulting insolubilised complex, and checking for the presence of the label attached to the anti-antigen (Angi-Ag₁) or anti-antibody (Anti-Ab₁).
Claim 15
Independent Claim
105 words
A method according to any one of claims 1 to 11 wherein the sample contains a hapten, the soluble complex is formed by reacting the sample in the liquid phase with a said matrix carrying ligand (X) and a sp ecific anti-hapten (Anti-H) in the presence of a labeled hapten probe (H-Z), allowing competitive binding to occur between the hapten (H) in the sample and the labeled hapten probe (H-Z) for anti-hapten (Anti-H) binding sites on the matrix, reacting the resulting soluble complex with immobilized anti-ligand (Y) on a solid support to form an insolubilised complex, washing insolubilised complex and checking for the label (Z).
Claim 16
Independent Claim
101 words
A method of measuring the level of a hapten in a sample, which comprises reacting the sample with a ligand-labeled specific antibody in the presence of a hapten labeled probe, allowing competitive binding to occur between the hapten in the sample and the labeled hapten probe for binding sites on the ligand-labeled antibody, adding an anti-ligand immobilized on a solid support directed against the ligand attached to the specific antibody, washing the solid phase, and checking for the presence of the label of the labeled hapten probe to provide a measure inversely proportional to the concentration of hapten in the sample.
Claim 17
Independent Claim
101 words
A method of measuring the level of a hapten in a sample, which comprises reacting the sample with a ligand-labeled specific antibody in the presence of a hapten labeled probe, allowing competitive binding to occur between the hapten in the sample and the labeled hapten probe for binding sites on the ligand-labeled antibody, adding an anti-ligand immobilized on a solid support directed against the ligand attached to the specific antibody, washing the solid phase, and checking for the presence of the label of the labeled hapten probe to provide a measure inversely proportional to the concentration of hapten in the sample.
References Cited (Backward Citations)
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