EP0253464A1
Expired / Lapsed
EP
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Methods for providing internal references for use in analyte receptor assays
Assignee: Hybritech Inc
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Filing: 1987-03-20
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Granted: —
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Expires: 2007-03-20
Abstract
Abstract A method is disclosed for providing internal reference for use in an analyte-receptor assay, for the determination of a target analyte in a sample. The assay uses a signal producing system and an analyte receptor capable of binding the target analyte at a discrete test zone on a solid phase, in an amount directly proportional to the amount of target analyte in the sample. The method comprises binding a reference receptor, selected to bind with an analyte receptor conjugate capable of complexation with the target analyte, at a discrete reference zone on a solid phase. The reference receptor is selected such that the rates of binding of the analyte receptor conjugate to the reference receptor and to the target analyte are directly proportional to the amounts of reference receptor at the reference zone and target analyte bound at the test zone on the solid phase, and the rate constants are substantially equivalent.
Inventors
- 👤 Gunars Edward Valkirs
- 👤 Richard Ray Anderson
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 17:30
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54366; G01N33/54386
Claims Analysis
Total Claims: 39
Claim 1
Independent Claim
175 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of a target analyte in a sample, the assay employing a signal producing system and an analyte receptor capable of binding the target analyte in an amount directly proportional to the amount of the target analyte in the sample, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptor is bound, a reference receptor selected to bind with an analyte receptor conjugate which is capable of complexation with the target analyte such that the rates of binding of the analyte receptor conjugate to the reference receptor and to the target analyte are directly proportional to the amounts of the reference receptor and the target analyte bound to the solid phase; the rate constants are substantially equivalent; and whereby signal development at the reference zone and the test zone is directly proportional to the amount of the analyte receptor conjugate bound to the reference and test zones.
Claim 2
Independent Claim
175 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of a target analyte in a sample, the assay employing a signal producing system and an analyte receptor capable of binding the target analyte in an amount directly proportional to the amount of the target analyte in the sample, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptor is bound, a reference receptor selected to bind with an analyte receptor conjugate which is capable of complexation with the target analyte such that the rates of binding of the analyte receptor conjugate to the reference receptor and to the target analyte are directly proportional to the amounts of the reference receptor and the target analyte bound to the solid phase; the rate constants are substantially equivalent; and whereby signal development at the reference zone and the test zone is directly proportional to the amount of the analyte receptor conjugate bound to the reference and test zones.
Claim 3
Dependent Claim
28 words
A method according to Claim 1 in which said reference receptor is a target analyte, an anti-enzyme antibody, an anti-hapten antibody, an anti-antibody antibody or an anti-idiotype antibody.
Claim 4
Dependent Claim
26 words
A method as claimed in Claim 2 in which the anti-enzyme antibody, said anti-hapten antibody, said anti-antibody antibody or said anti-idiotype antibody is a monoclonal antibody.
Claim 5
Dependent Claim
21 words
A method as claimed in Claim 2 in which the anti-anti-body or the anti-idiotype antibody are specific for a monoclonal antibody.
Claim 6
Dependent Claim
15 words
A method according to Claim 1 in which said analyte receptor is a monoclonal antibody.
Claim 7
Dependent Claim
32 words
A method as claimed in Claim 1 in which the analyte receptor is a monoclonal antibody and the target analyte is HCG, LH, FSH, CKMB, CKMM, AFP or hepatitis B surface antigen.
Claim 8
Dependent Claim
22 words
A method as claimed in Claim 1 in which the analyte receptor is an allergen and the target analyte is allergen-specific IgE.
Claim 9
Independent Claim
191 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of a target analyte in a sample, said assay employing a signal producing system and an analyte receptor capable of binding the target analyte in an amount directly proportional to the amount of the target analyte in the sample, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptor is bound, a ligand conjugate reference receptor selected to bind with a ligand conjugate, such that the rate of binding of the ligand conjugate to the reference receptor and the rate of binding of an analyte receptor conjugate to the target analyte are directly proportional to the amount of the reference receptor and the target analyte bound to the solid phase; the rate constants are substantially equivalent; and whereby signal development at the reference zone is directly proportional to the amount of the ligand conjugate bound to the reference zone and signal development at said test zone is directly proportional to the amount of the analyte receptor conjugate bound to said test zone.
Claim 10
Independent Claim
191 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of a target analyte in a sample, said assay employing a signal producing system and an analyte receptor capable of binding the target analyte in an amount directly proportional to the amount of the target analyte in the sample, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptor is bound, a ligand conjugate reference receptor selected to bind with a ligand conjugate, such that the rate of binding of the ligand conjugate to the reference receptor and the rate of binding of an analyte receptor conjugate to the target analyte are directly proportional to the amount of the reference receptor and the target analyte bound to the solid phase; the rate constants are substantially equivalent; and whereby signal development at the reference zone is directly proportional to the amount of the ligand conjugate bound to the reference zone and signal development at said test zone is directly proportional to the amount of the analyte receptor conjugate bound to said test zone.
Claim 11
Dependent Claim
18 words
A method as claimed in Claim 8 in which the ligand is a protein, enzyme, hapten or oligonucleotide.
Claim 12
Dependent Claim
26 words
A method as claimed in claim 9 in which the ligand is hen egg-white lysozyme, bovine serum albumin, horseradish peroxidase, bromelin, fluorescein or a metal chelator.
Claim 13
Dependent Claim
19 words
A method as claimed in claim 8 in which the reference receptor and said analyte receptor are monoclonal antibodies.
Claim 14
Dependent Claim
21 words
A method according to claim 1 or 8 in which the label component of the analyte receptor conjugate is an enzyme.
Claim 15
Dependent Claim
17 words
A method as claimed in Claim 12 in which the enzyme is alkaline phosphatase or horseradish peroxidase.
Claim 16
Dependent Claim
196 words
An analyte-receptor assay process for the determination of a target analyte in a sample which comprises: a) introducing the sample suspected of containing the target analyte onto a solid phase comprising a discrete test zone to which an analyte receptor capable of binding with the target analyte is bound, the solid phase further comprising at least one discrete reference zone to which a reference receptor is bound, the analyte receptor being bound in a manner such that a direct proportion of the amount of the target analyte in the sample binds to the test zone; b) adding a solution of analyte receptor conjugate, the analyte receptor conjugate being labeled to permit detection by signal development, in order to bind the analyte receptor conjugate to the target analyte and to the reference receptor such that the rates of binding of the analyte receptor conjugate to the test zone and to said reference zone are directly proportional to the amounts of the target analyte and the reference receptor bound to the solid phase and the rate constants are substantially equivalent; and c) determining signal development generated by the analyte receptor conjugate bound to the test and reference zones.
Claim 17
Dependent Claim
196 words
An analyte-receptor assay process for the determination of a target analyte in a sample which comprises: a) introducing the sample suspected of containing the target analyte onto a solid phase comprising a discrete test zone to which an analyte receptor capable of binding with the target analyte is bound, the solid phase further comprising at least one discrete reference zone to which a reference receptor is bound, the analyte receptor being bound in a manner such that a direct proportion of the amount of the target analyte in the sample binds to the test zone; b) adding a solution of analyte receptor conjugate, the analyte receptor conjugate being labeled to permit detection by signal development, in order to bind the analyte receptor conjugate to the target analyte and to the reference receptor such that the rates of binding of the analyte receptor conjugate to the test zone and to said reference zone are directly proportional to the amounts of the target analyte and the reference receptor bound to the solid phase and the rate constants are substantially equivalent; and c) determining signal development generated by the analyte receptor conjugate bound to the test and reference zones.
Claim 18
Dependent Claim
213 words
An analyte-receptor assay process for the determination of a target analyte in a sample comprising: a) introducing the sample suspected of containing the target analyte onto a solid phase comprising a discrete test zone to which an analyte receptor capable of binding the target analyte is bound, the solid phase further comprising at least one discrete reference zone to which a ligand conjugate reference receptor is bound, the analyte receptor being bound such that a direct proportion of the amount of said target analyte in said sample binds to said test zone; b) adding a solution comprising analyte receptor conjugate and ligand conjugate, the analyte receptor conjugate and the ligand conjugate being labeled to permit detection by signal development, in order to bind said analyte receptor conjugate to said target analyte and said ligand conjugate to the reference receptor such that the rates of binding of the analyte receptor conjugate to the test zone and said ligand conjugate to the reference zone are directly proportional to the amounts of the target analyte and the reference receptor bound to the solid phase and the rate constants are substantially equivalent; and c) determining signal development generated by the analyte receptor conjugate bound to the test zone and the ligand conjugate bound to the reference zone.
Claim 19
Dependent Claim
213 words
An analyte-receptor assay process for the determination of a target analyte in a sample comprising: a) introducing the sample suspected of containing the target analyte onto a solid phase comprising a discrete test zone to which an analyte receptor capable of binding the target analyte is bound, the solid phase further comprising at least one discrete reference zone to which a ligand conjugate reference receptor is bound, the analyte receptor being bound such that a direct proportion of the amount of said target analyte in said sample binds to said test zone; b) adding a solution comprising analyte receptor conjugate and ligand conjugate, the analyte receptor conjugate and the ligand conjugate being labeled to permit detection by signal development, in order to bind said analyte receptor conjugate to said target analyte and said ligand conjugate to the reference receptor such that the rates of binding of the analyte receptor conjugate to the test zone and said ligand conjugate to the reference zone are directly proportional to the amounts of the target analyte and the reference receptor bound to the solid phase and the rate constants are substantially equivalent; and c) determining signal development generated by the analyte receptor conjugate bound to the test zone and the ligand conjugate bound to the reference zone.
Claim 20
Dependent Claim
22 words
A method as claimed in Claim 1 or 8 in which at least two reference receptors are bound to said solid phase.
Claim 21
Dependent Claim
20 words
A process as claimed in Claim 14 or 15 in which the solid phase comprises at least two reference receptors.
Claim 22
Independent Claim
186 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of multiple target analytes in a sample wherein the target analytes have a common site to which an analyte receptor conjugate is capable of binding, and wherein the assay employs a signal producing system and analyte receptors capable of binding the target analytes in amounts directly proportional to the amounts of the target analytes in the sample, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptors are bound, at least one reference receptor selected to bind with the analyte receptor conjugate such that the rates of binding of the analyte receptor conjugate to the reference receptor and to the target analytes are directly proportional to the amounts of the reference receptor and the target analytes bound to the solid phase; the rate constants are substantially equivalent; and whereby signal development at the reference zone and the test zones is directly proportional to the amounts of the analyte receptor conjugate bound to the reference and test zones.
Claim 23
Independent Claim
186 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of multiple target analytes in a sample wherein the target analytes have a common site to which an analyte receptor conjugate is capable of binding, and wherein the assay employs a signal producing system and analyte receptors capable of binding the target analytes in amounts directly proportional to the amounts of the target analytes in the sample, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptors are bound, at least one reference receptor selected to bind with the analyte receptor conjugate such that the rates of binding of the analyte receptor conjugate to the reference receptor and to the target analytes are directly proportional to the amounts of the reference receptor and the target analytes bound to the solid phase; the rate constants are substantially equivalent; and whereby signal development at the reference zone and the test zones is directly proportional to the amounts of the analyte receptor conjugate bound to the reference and test zones.
Claim 24
Dependent Claim
20 words
A method as claimed in Claim 18 in which the target analytes are FSH and LH, or CKMB and CKMM.
Claim 25
Independent Claim
232 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of multiple target analytes in a sample wherein the assay employs a signal producing system, a ligand conjugate, analyte receptors capable of binding said target analytes in amounts directly proportional to the amounts of the target analytes in said sample, and analyte receptor conjugates in a number equal to the number of target analytes to be determined, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptors are bound, a ligand conjugate reference receptor selected to bind with the ligand conjugate, said ligand conjugate and said analyte receptor conjugates being further selected such that the rates of binding of the ligand conjugate to the reference receptor and the analyte receptor conjugates to the target analytes are directly proportional to the amounts of the reference receptor and the target analytes bound to the solid phase, the rate constants for binding of the ligand conjugate to said reference receptor and for binding of said analyte receptor conjugates to the target analytes are substantially equivalent; and whereby signal development at the reference zone and the test zones are directly proportional to the amount of the ligand conjugate bound to the reference zone and to the amounts of the analyte receptor conjugates bound to the test zones.
Claim 26
Independent Claim
232 words
A method for providing internal references for use in a sequential analyte-receptor assay for the determination of multiple target analytes in a sample wherein the assay employs a signal producing system, a ligand conjugate, analyte receptors capable of binding said target analytes in amounts directly proportional to the amounts of the target analytes in said sample, and analyte receptor conjugates in a number equal to the number of target analytes to be determined, which comprises: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptors are bound, a ligand conjugate reference receptor selected to bind with the ligand conjugate, said ligand conjugate and said analyte receptor conjugates being further selected such that the rates of binding of the ligand conjugate to the reference receptor and the analyte receptor conjugates to the target analytes are directly proportional to the amounts of the reference receptor and the target analytes bound to the solid phase, the rate constants for binding of the ligand conjugate to said reference receptor and for binding of said analyte receptor conjugates to the target analytes are substantially equivalent; and whereby signal development at the reference zone and the test zones are directly proportional to the amount of the ligand conjugate bound to the reference zone and to the amounts of the analyte receptor conjugates bound to the test zones.
Claim 27
Dependent Claim
19 words
A method according to Claim 20 in which said target analytes are FSH and LH or CKMB and CKMM.
Claim 28
Independent Claim
227 words
A method for providing internal references for use in an analyte-receptor assay for the determination of a target analyte in a sample wherein said assay employs an analyte-receptor conjugate, said analyte receptor conjugate being pre-complexed with said target analyte to form a target analyte complex in an amount directly proportional to the amount of said target analyte in said sample, an analyte receptor capable of binding said target analyte complex in an amount directly proportional to the amount of said target analyte complex, a ligand conjugate and a signal producing system, said method comprising: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which analyte receptor is bound, a ligand conjugate reference receptor selected to bind with a ligand conjugate, said reference receptor and said analyte receptor being further selected such that the rate of binding for said ligand conjugate to said reference receptor and the rate of binding for said target analyte complex to said analyte receptor are directly proportional to the amounts of said ligand conjugate and said target analyte complex; and whereby signal development at the reference zone is directly proportional to the amount of the ligand conjugate bound to the reference zone and signal development at the test zone is directly proportional to the amount of the target analyte complex bound to the test zone.
Claim 29
Independent Claim
227 words
A method for providing internal references for use in an analyte-receptor assay for the determination of a target analyte in a sample wherein said assay employs an analyte-receptor conjugate, said analyte receptor conjugate being pre-complexed with said target analyte to form a target analyte complex in an amount directly proportional to the amount of said target analyte in said sample, an analyte receptor capable of binding said target analyte complex in an amount directly proportional to the amount of said target analyte complex, a ligand conjugate and a signal producing system, said method comprising: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which analyte receptor is bound, a ligand conjugate reference receptor selected to bind with a ligand conjugate, said reference receptor and said analyte receptor being further selected such that the rate of binding for said ligand conjugate to said reference receptor and the rate of binding for said target analyte complex to said analyte receptor are directly proportional to the amounts of said ligand conjugate and said target analyte complex; and whereby signal development at the reference zone is directly proportional to the amount of the ligand conjugate bound to the reference zone and signal development at the test zone is directly proportional to the amount of the target analyte complex bound to the test zone.
Claim 30
Dependent Claim
26 words
A method according to Claim 22 in which said ligand is selected from hen egg-white lysozyme, bovine serum albumin, horseradish peroxidase, bromelin, fluorescein and metal chelators.
Claim 31
Dependent Claim
40 words
A method according to Claim 22 wherein the label component of said analyte receptor conjugate and said ligand conjugate is selected from an enzyme, a radionuclide, a fluorescent agent, a phosphorescent agent, or a polymer containing dyes or chemiluminescent moieties.
Claim 32
Dependent Claim
12 words
A method according to Claim 24 wherein said label is an enzyme.
Claim 33
Dependent Claim
15 words
A method according to Claim 25 wherein said enzyme is alkaline phosphatase or horseradish peroxidase.
Claim 34
Independent Claim
233 words
A method for providing internal references for use in an analyte-receptor assay for the determination of multiple target analytes in a sample wherein said assay employs analyte-receptor conjugates in a number equal to the number of target analytes to be determined, the analyte receptor conjugates being pre-complexed with the target analytes to form target analyte complexes in amounts directly proportional to the amounts of the target analytes in the sample, analyte receptors capable of binding the target analyte complexes in amounts directly proportional to the amounts of said target analyte complexes, a ligand conjugate and a signal producing system, said method comprising: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptors are bound, a ligand conjugate reference receptor selected to bind the ligand conjugate, said reference receptor and said analyte receptors being further selected such that the rate of binding the ligand conjugate to the reference receptor and the rates of binding the target analyte complexes to the analyte receptors are directly proportional to the amounts of the ligand conjugate and the target analyte complexes; and whereby signal development at the reference zone is directly proportional to the amount of the ligand conjugate bound to said reference zone and signal development at the test zones is directly proportional to the amounts of the target analyte complexes bound to the test zones.
Claim 35
Independent Claim
233 words
A method for providing internal references for use in an analyte-receptor assay for the determination of multiple target analytes in a sample wherein said assay employs analyte-receptor conjugates in a number equal to the number of target analytes to be determined, the analyte receptor conjugates being pre-complexed with the target analytes to form target analyte complexes in amounts directly proportional to the amounts of the target analytes in the sample, analyte receptors capable of binding the target analyte complexes in amounts directly proportional to the amounts of said target analyte complexes, a ligand conjugate and a signal producing system, said method comprising: binding to a solid phase, at a discrete reference zone separate from a discrete test zone to which the analyte receptors are bound, a ligand conjugate reference receptor selected to bind the ligand conjugate, said reference receptor and said analyte receptors being further selected such that the rate of binding the ligand conjugate to the reference receptor and the rates of binding the target analyte complexes to the analyte receptors are directly proportional to the amounts of the ligand conjugate and the target analyte complexes; and whereby signal development at the reference zone is directly proportional to the amount of the ligand conjugate bound to said reference zone and signal development at the test zones is directly proportional to the amounts of the target analyte complexes bound to the test zones.
Claim 36
Independent Claim
159 words
An apparatus for use in an analyte-receptor assay for the determination of a target analyte in a sample comprising: a) a porous solid phase member comprising a discrete test zone to which is bound analyte receptor capable of binding said target analyte, and at least one discrete reference zone to which is bound reference receptor, said reference receptor being selected to bind with an analyte receptor conjugate capable of binding with said target analyte, said reference receptor being further selected such that the rates for binding of said analyte receptor conjugate to said reference receptor and to said target analyte are directly proportional to the amounts of said reference receptor and said target analyte bound to said solid phase, and such that the rate constants are substantially equivalent, whereby signal development at said reference zone and said test zone is directly proportional to the amount of said analyte receptor conjugate bound to said reference zone and said test zone.
Claim 37
Independent Claim
159 words
An apparatus for use in an analyte-receptor assay for the determination of a target analyte in a sample comprising: a) a porous solid phase member comprising a discrete test zone to which is bound analyte receptor capable of binding said target analyte, and at least one discrete reference zone to which is bound reference receptor, said reference receptor being selected to bind with an analyte receptor conjugate capable of binding with said target analyte, said reference receptor being further selected such that the rates for binding of said analyte receptor conjugate to said reference receptor and to said target analyte are directly proportional to the amounts of said reference receptor and said target analyte bound to said solid phase, and such that the rate constants are substantially equivalent, whereby signal development at said reference zone and said test zone is directly proportional to the amount of said analyte receptor conjugate bound to said reference zone and said test zone.
Claim 38
Dependent Claim
38 words
An apparatus according to Claim 28 wherein said solid phase member comprises a porous matrix in which analyte receptor-bound microspheres are entrapped within said discrete test zone and reference receptor-bound microspheres are entrapped within said discrete reference zone.
Claim 39
Dependent Claim
37 words
An apparatus as claimed in Claim 28 or 29 comprising a means, operatively associated with said porous solid phase member, for facilitating the flow of said fluid sample and assay reagents through said porous solid phase member.
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