US4737456A
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US
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Reducing interference in ligand-receptor binding assays
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1985-05-09
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Granted: 1988-04-12
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Expires: 2005-05-09
Abstract
Abstract A method for assaying for a ligand analyte which is a member of a specific binding pair ("sbp member") consisting of ligand and its complementary receptor is disclosed. The ligand analyte has a binding site in common with an interfering substance present in a sample suspected of containing the analyte. The interfering substance has at least two binding sites. The method comprises combining in an assay medium without intervening separation (1) the sample, (2) a blocking receptor which does not bind to the ligand and does bind to the interfering substance, thereby blocking the binding of a common receptor to the interfering substance, and (3) a common receptor which binds to the common binding site. Any additional members of a signal producing system capable of producing a detectable signal in relation to the amount of analyte in the sample are added to the assay medium. Next, the assay medium is examined for the presence of a detectable signal. The method has application to both heterogeneous and homogeneous assays.
Inventors
- 👤 Litai Weng
- 👤 Iam Gibbons
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 16:33
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/5306; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/962; Y10S435/975; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/815; Y10S436/817; Y10S436/818; Y10S436/819; Y10S436/825
Claims Analysis
Total Claims: 41
Claim 1
Independent Claim
141 words
A method for assaying for a ligand which is a member of a specific binding pair ("sbp member ") consisting of said ligand and its complementary receptor wherein said ligand has a binding site in common with an interfering substance present in a sample suspected of containing said ligand, said interfering substance having at least two binding sites, which method comprises combining without intervening separation (1) said sample with (2) a blocking receptor, which does not bind to said ligand and does bind to said interfering substance and thereby blocks binding of a common receptor to said interfering substance, and (3) a common receptor which binds to said common binding site, to provide an assay medium, and determining the extent of binding of said common receptor, the extent of binding being related to the amount of said analyte in said sample.
Claim 2
Independent Claim
141 words
A method for assaying for a ligand which is a member of a specific binding pair ("sbp member ") consisting of said ligand and its complementary receptor wherein said ligand has a binding site in common with an interfering substance present in a sample suspected of containing said ligand, said interfering substance having at least two binding sites, which method comprises combining without intervening separation (1) said sample with (2) a blocking receptor, which does not bind to said ligand and does bind to said interfering substance and thereby blocks binding of a common receptor to said interfering substance, and (3) a common receptor which binds to said common binding site, to provide an assay medium, and determining the extent of binding of said common receptor, the extent of binding being related to the amount of said analyte in said sample.
Claim 3
Dependent Claim
12 words
The method of claim 1 wherein said common receptor is an antibody.
Claim 4
Dependent Claim
13 words
The method of claim 1 wherein said blocking receptor is a monoclonal antibody.
Claim 5
Dependent Claim
11 words
The method of claim 1 wherein said ligand is a protein.
Claim 6
Dependent Claim
13 words
The method of claim 1 wherein said blocking and common receptors are antibodies.
Claim 7
Dependent Claim
23 words
The method of claim 1 wherein said ligand is human thyroid stimulating hormone (hTSH) and said interfering substance is human chorionic gonadotropin (hCG).
Claim 8
Dependent Claim
29 words
The method of claim 6 wherein said common receptor recognizes the alpha sub-unit of hTSH and hCG and said blocking receptor is specific for the beta sub-unit of hCG.
Claim 9
Dependent Claim
12 words
The method of claim 1 which is an enzyme linked immunosorbent assay.
Claim 10
Dependent Claim
11 words
The method of claim 1 wherein said common receptor is immobilized.
Claim 11
Dependent Claim
14 words
The method of claim 1 wherein said common receptor is conjugated to a label.
Claim 12
Dependent Claim
11 words
The method of claim 10 wherein said label is an enzyme.
Claim 13
Dependent Claim
11 words
The method of claim 10 wherein said label is a radioisotope.
Claim 14
Dependent Claim
33 words
The method of claim 10 wherein said label is part of a signal producing system and said determining is carried out by examining said assay medium for the presence of a detectible signal.
Claim 15
Dependent Claim
104 words
In an assay method for a ligand which is a member of a specific binding pair ("sbp member") consisting of said ligand and its complementary receptor wherein said ligand has a binding site in common with a binding site of an interfering substance having at least two binding sites, said method employing a receptor recognizing the common binding site, the improvement which comprises adding to an assay medium containing said analyte and said interfering substance, without intervening separation, a specific binding compound which blocks the binding between said receptor and said interfering substance and does not block the binding between said receptor and ligand.
Claim 16
Dependent Claim
104 words
In an assay method for a ligand which is a member of a specific binding pair ("sbp member") consisting of said ligand and its complementary receptor wherein said ligand has a binding site in common with a binding site of an interfering substance having at least two binding sites, said method employing a receptor recognizing the common binding site, the improvement which comprises adding to an assay medium containing said analyte and said interfering substance, without intervening separation, a specific binding compound which blocks the binding between said receptor and said interfering substance and does not block the binding between said receptor and ligand.
Claim 17
Dependent Claim
11 words
The method of claim 14 wherein said receptor is an antibody.
Claim 18
Dependent Claim
12 words
The method of claim 14 wherein said receptor is a monoclonal antibody.
Claim 19
Dependent Claim
11 words
The method of claim 14 wherein said ligand is a protein.
Claim 20
Dependent Claim
17 words
The method of claim 14 wherein said compound is an antibody that binds to said interfering substance.
Claim 21
Dependent Claim
117 words
In an assay method for an antigen which is a member of a specific binding pair ("sbp member") consisting of said antigen and its complementary antibody wherein said antigen has at least two epitopic sites, one of said epitopic sites being common to an interfering substance having at least two epitopic sites, said method employing a common antibody, said common antibody recognizing the common epitopic site, the improvement which comprises combining with an assay medium containing said antigen and said interfering substance an amount of a blocking antibody sufficient to block the binding between said common antibody and said intefering substance wherein said blocking antibody does not block the binding between said common antibody and said antigen.
Claim 22
Dependent Claim
117 words
In an assay method for an antigen which is a member of a specific binding pair ("sbp member") consisting of said antigen and its complementary antibody wherein said antigen has at least two epitopic sites, one of said epitopic sites being common to an interfering substance having at least two epitopic sites, said method employing a common antibody, said common antibody recognizing the common epitopic site, the improvement which comprises combining with an assay medium containing said antigen and said interfering substance an amount of a blocking antibody sufficient to block the binding between said common antibody and said intefering substance wherein said blocking antibody does not block the binding between said common antibody and said antigen.
Claim 23
Dependent Claim
13 words
The method of claim 19 wherein said common antibody is a monoclonal antibody.
Claim 24
Dependent Claim
11 words
The method of claim 19 wherein said antigen is a protein.
Claim 25
Dependent Claim
23 words
The method of claim 19 wherein said antigen is human thyroid stimulating hormone (hTSH) and said interfering substance is human chorionic gonadotropin (hCG).
Claim 26
Dependent Claim
34 words
The method of claim 22 wherein said common antibody recognizes the alph sub-unit of TSH and HCG and said blocking antibody binds only to intact HCG comprised of both the alpha and beta sub-units.
Claim 27
Dependent Claim
12 words
The method of claim 19 which is an enzyme linked immunosorbent assay.
Claim 28
Dependent Claim
11 words
The method of claim 19 wherein said common antibody is immobilized.
Claim 29
Dependent Claim
111 words
In an assay method for a protein analyte having alpha and beta sub-units, wherein one of said sub-units is common to a sub-unit of an interfering protein having alpha and beta sub-units, said method employing a common antibody which recognizes the sub-unit common to said protein analyte and said interfering substance and an antibody specific for the non-common sub-unit of said analyte, the improvement which comprises combinine with said assay medium containing said interfering substance an amount of a blocking antibody that binds specifically to said interfering subtance but not to its separate alpha and beta sub-units, said blocking antibody blocking the binding of said interfering substance to said common antibody.
Claim 30
Dependent Claim
111 words
In an assay method for a protein analyte having alpha and beta sub-units, wherein one of said sub-units is common to a sub-unit of an interfering protein having alpha and beta sub-units, said method employing a common antibody which recognizes the sub-unit common to said protein analyte and said interfering substance and an antibody specific for the non-common sub-unit of said analyte, the improvement which comprises combinine with said assay medium containing said interfering substance an amount of a blocking antibody that binds specifically to said interfering subtance but not to its separate alpha and beta sub-units, said blocking antibody blocking the binding of said interfering substance to said common antibody.
Claim 31
Dependent Claim
23 words
The method of claim 6 wherein said protein analyte is human thyroid stimulating hormone (hTSH) and said interfering substance is human chorionic gonadotropin.
Claim 32
Dependent Claim
93 words
A composition comprising one or both of (1) a common receptor that specifically binds to a binding site common to an endogenous interfering substance and a ligand suspected of being present in a sample and (2) members of a signal producing system capable of producing a detectible signal in the presence of said ligand in combination with (3) an amount of a blocking receptor which does not bind to said ligand and does bind to said endogenous interfering substance and thereby blocks the binding between said common receptor and said endogenous interfering substance.
Claim 33
Dependent Claim
93 words
A composition comprising one or both of (1) a common receptor that specifically binds to a binding site common to an endogenous interfering substance and a ligand suspected of being present in a sample and (2) members of a signal producing system capable of producing a detectible signal in the presence of said ligand in combination with (3) an amount of a blocking receptor which does not bind to said ligand and does bind to said endogenous interfering substance and thereby blocks the binding between said common receptor and said endogenous interfering substance.
Claim 34
Dependent Claim
12 words
The composition of claim 28 wherein said blocking receptor is an antibody.
Claim 35
Dependent Claim
13 words
The composition of claim 28 wherein said common receptor is a monoclonal antibody.
Claim 36
Dependent Claim
11 words
The composition of claim 28 wherein said ligand is a protein.
Claim 37
Dependent Claim
13 words
The composition of claim 28 wherein said blocking and common receptors are antibodies.
Claim 38
Dependent Claim
23 words
The composition of claim 28 wherein said ligand is human thyroid stimulating hormone (hTSH) and said interfering substance is human chorionic gonadotropin (hCG).
Claim 39
Dependent Claim
35 words
The composition of claim 33 wherein said common receptor recognizes the alpha sub-unit of hTSH and hCG and said blocking receptor binds only to intact hCG comprised of both alpha and beta sub-units of hCG.
Claim 40
Dependent Claim
18 words
The composition of claim 28 which additionally comprises a third receptor specific for non-common site on said ligand.
Claim 41
Dependent Claim
22 words
A kit for use in an assay method, comprising in packaged combination the composition of claim 28, and ancillary reagents as required.
References Cited (Backward Citations)
US3935074A
Cited
US4376110A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-04-12 | 1991-10-12 | 1992-04-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-04-12 | 1995-10-12 | 1996-04-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-04-12 | 1999-10-12 | 2000-04-12 | $7700.00 | Window Closed / Lapsed |