US4235869A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Assay employing a labeled Fab-fragment ligand complex
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1978-05-16
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Granted: 1980-11-25
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Expires: 1998-05-16
Abstract
Abstract Methods and compositions are provided for improved protein binding assays by preparing compositions having indirectly labeled ligands substantially free of label conjugated to materials other than the indirectly labeled ligand. The method for preparing the compositions involves employing a monovalent receptor to which the label is conjugated and combining the monovalent receptor labeled conjugate to the ligand, either in pure or impure form. The mixture is then segregated according to molecular weight and the ligand conjugated to the labeled receptor isolated. This conjugate may then be directly used as a reagent in a protein binding assay, where the assay mixture is substantially free of label other than labeled receptor bound to ligand. The labels will be for the most part of relatively low molecular weight, while the receptor is preferably a Fab fragment. The ratio of receptor to ligand will be chosen so as to provide reasonable molecular weight distinctions between unbound ligand, unbound labeled receptor, ligand bound to labeled receptor, and other materials which may be in the mixture. Various techniques may be used for the separation. The assays are performed in accordance with known methods employing a second receptor composition, which may be labeled or unlabeled.
Inventors
- 👤 Moshe Schwarzberg
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 18:57
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/563; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S436/00; Y10S436/80
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
171 words
A method for performing a protein binding assay employing a labeled ligand complex reagent, where the ligand is a protein and a member of a specific binding pair consisting of ligand and antiligand and is polyepitopic and of at least about 15,000 molecular weight; and said label is of from about 100 to 2,000 molecular weight and provides a detectible signal affected by the proximity of antiligand due to the steric bulk of said antiligand or a group conjugated to said antiligand which interacts with said label; said labeled ligand complex reagent prepared by the method comprising: (a) combining said ligand with a γ-globulin composition containing Fab fragments derived from antiligand, said R-globulin composition having been conjugated with said label, wherein said ligand and labeled fab fragments form complexes having at least one free ligand epitopic site; and (b) separating said complexes from other labeled protein; said protein binding assay comprising; combining said complexes with a sample suspected of containing said ligand in an assay medium for determination of said ligand.
Claim 2
Independent Claim
171 words
A method for performing a protein binding assay employing a labeled ligand complex reagent, where the ligand is a protein and a member of a specific binding pair consisting of ligand and antiligand and is polyepitopic and of at least about 15,000 molecular weight; and said label is of from about 100 to 2,000 molecular weight and provides a detectible signal affected by the proximity of antiligand due to the steric bulk of said antiligand or a group conjugated to said antiligand which interacts with said label; said labeled ligand complex reagent prepared by the method comprising: (a) combining said ligand with a γ-globulin composition containing Fab fragments derived from antiligand, said R-globulin composition having been conjugated with said label, wherein said ligand and labeled fab fragments form complexes having at least one free ligand epitopic site; and (b) separating said complexes from other labeled protein; said protein binding assay comprising; combining said complexes with a sample suspected of containing said ligand in an assay medium for determination of said ligand.
Claim 3
Dependent Claim
12 words
A method according to claim 1 wherein said label is a fluorescer.
Claim 4
Dependent Claim
13 words
A method according to claim 1, wherein said protein ligand is a globulin.
Claim 5
Dependent Claim
12 words
A method according to claim 3 wherein said label is a chemiluminescer.
Claim 6
Independent Claim
150 words
A method for performing a protein binding assay for immunoglobulin employing a labeled immunoglobulin complex reagent wherein said label is of from about 100 to 2,000 molecular weight and provides a detectible signal affected by the proximity of antiimunoglobulin due to the steric bulk of said antiimmunoglobulin or a group conjugated to said antiimmunoglobulin which interacts with said label; said labeled immunoglobulin complex reagent prepared by the method comprising: (a) combining said immunoglobulin with a γ-globulin composition containing Fab fragments derived from a serum protein composition having antiimunoglobulin and other protein conjugated with said label, wherein said Fab fragments derived from antiimunoglobulin and said immunoglobulin form complexes having at least one free ligand epitopic site; and (b) separating said complexes from the other labeled protein; said protein binding assay comprising: combining said complexes with a sample suspected of containing said immunoglobulin in an assay medium for determination of said immunoglobulin.
Claim 7
Independent Claim
150 words
A method for performing a protein binding assay for immunoglobulin employing a labeled immunoglobulin complex reagent wherein said label is of from about 100 to 2,000 molecular weight and provides a detectible signal affected by the proximity of antiimunoglobulin due to the steric bulk of said antiimmunoglobulin or a group conjugated to said antiimmunoglobulin which interacts with said label; said labeled immunoglobulin complex reagent prepared by the method comprising: (a) combining said immunoglobulin with a γ-globulin composition containing Fab fragments derived from a serum protein composition having antiimunoglobulin and other protein conjugated with said label, wherein said Fab fragments derived from antiimunoglobulin and said immunoglobulin form complexes having at least one free ligand epitopic site; and (b) separating said complexes from the other labeled protein; said protein binding assay comprising: combining said complexes with a sample suspected of containing said immunoglobulin in an assay medium for determination of said immunoglobulin.
Claim 8
Dependent Claim
11 words
A method according to claim 5, wherein said immunoglobulin is IgG.
Claim 9
Dependent Claim
11 words
A method according to claim 5, wherein said immunoglobulin is IgM.
Claim 10
Dependent Claim
11 words
A method according to claim 5, wherein said immunoglobulin is IgA.
Claim 11
Dependent Claim
10 words
A method according to claim 5, wherein said immunoglobulin IgE.
Claim 12
Independent Claim
17 words
A method according to any one of claims 5 to 9, wherein said label is a fluorescer.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-11-25 | 1984-05-26 | 1984-11-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-11-25 | 1988-05-26 | 1988-11-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-11-25 | 1992-05-26 | 1992-11-25 | $7700.00 | Window Closed / Lapsed |