Patent US439088A successfully refreshed from Google Patents.
US5045480A
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Gel particles having hapten moieties bound thereto as immunoassay reagent
Assignee: Bayer Corp
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Filing: 1989-02-09
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Granted: 1991-09-03
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Expires: 2009-02-09
Abstract
Abstract An immobilized hapten reagent for use in a specific binding assay for the determination of a hapten or binding analog thereof in a liquid test sample and methods for preparing and using the immobilized hapten reagent. The immobilized hapten reagent comprises a hapten moiety covalenty linked substantially only to the external surface of a gel particle. The immobilized hapten reagent is prepared by forming a reaction mixture comprising the hapten moiety and the carrier material in a nonswelling solvent wherein the gel particle is substantially impervious to the hapten moiety and wherein an analytically insignificant amount of the hapten moiety is nonspecifically bound to the gel particle. The immobilized hapten reagent is characterized by being substantially stable in aqueous solutions and exhibits an insignificant amount of leakage of the hapten moiety into a liquid test medium.
Inventors
- 👤 Richard D. Johnson
- 👤 H.-Volker Runzheimer
- 👤 Ronald G. Sommer
- 👤 Kin F. Yip
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 05:15
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54353; G01N33/54313
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
168 words
A method of preparing a stable immobilized hapten reagent for use in the specific binding assay determination of a hapten or binding analog thereof in a liquid test sample, the method comprising the steps of: (a) reacting a hapten moiety with a gel particle comprising a plurality of external and internal chemically active functional groups wherein the reaction is performed in a solvent in which the gel particle is substantially nonswollen and under conditions to form a covalent bond between the hapten moiety and said external active functional groups which is substantially stable in aqueous solutions; (b) washing the gel particle resulting from step(a) with a nonswelling solvent; (c) washing the gel particle resulting from step(b) with an aqueous solution; and (d) isolating the immobilized hapten reagent resulting from step(c) comprising said gel particle and said hapten moieties bound thereto wherein substantially all of said bound hapten moieties are covalently linked to the external surface functional groups by a linking group which is substantially stable in aqueous solutions.
Claim 2
Independent Claim
168 words
A method of preparing a stable immobilized hapten reagent for use in the specific binding assay determination of a hapten or binding analog thereof in a liquid test sample, the method comprising the steps of: (a) reacting a hapten moiety with a gel particle comprising a plurality of external and internal chemically active functional groups wherein the reaction is performed in a solvent in which the gel particle is substantially nonswollen and under conditions to form a covalent bond between the hapten moiety and said external active functional groups which is substantially stable in aqueous solutions; (b) washing the gel particle resulting from step(a) with a nonswelling solvent; (c) washing the gel particle resulting from step(b) with an aqueous solution; and (d) isolating the immobilized hapten reagent resulting from step(c) comprising said gel particle and said hapten moieties bound thereto wherein substantially all of said bound hapten moieties are covalently linked to the external surface functional groups by a linking group which is substantially stable in aqueous solutions.
Claim 3
Dependent Claim
31 words
The method of claim 1 wherein said gel particle is selected from the group consisting of polymers and copolymers of acrylic acid, agarose, cellulose, dextran, and their derivatives or mixtures thereof.
Claim 4
Dependent Claim
13 words
The method of claim 1 wherein the nonswelling solvent is an organic solvent.
Claim 5
Dependent Claim
26 words
The method of claim 3 wherein said organic solvent is selected from the group consisting of dimethylformamide, dimethylsulfoxide, acetone, chlorinated hydrocarbons, and cyclic and acyclic alkylethers.
Claim 6
Dependent Claim
22 words
The method of claim 1 wherein the hapten moiety is selected from the group consisting of drugs, metabolites, hormones, vitamins, and toxins.
Claim 7
Dependent Claim
22 words
The method of claim 1 wherein the hapten moiety is selected from the group consisting of digoxin, digitoxigenin, digitoxin, digoxigenin, and 12-0-acetyldigoxigenin.
Claim 8
Dependent Claim
14 words
The method of claim 1 wherein the hapten moiety is a glycosylated peptide sequence.
Claim 9
Dependent Claim
35 words
The method of claim 1 wherein less than from about 1×10 -12 moles hapten/gram gel particle dissociates from the gel particle of the immobilized hapten reagent upon standing in an aqueous solution for one week.
Claim 10
Dependent Claim
35 words
The method of claim 8 wherein less than from about 1×10 -13 moles hapten/gram gel particle dissociates from the gel particle of the immobilized hapten reagent upon standing in an aqueous solution for one week.
Claim 11
Dependent Claim
20 words
The method of claim 1 wherein the gel particle in its nonswollen state is substantially impervious to the hapten moiety.
Claim 12
Dependent Claim
17 words
The method of claim 1 wherein the gel particle is less than about 1,000 microns in diameter.
Claim 13
Dependent Claim
20 words
The method of claim 1 wherein the gel particle is from,-between, about 0.001 microns and about 850 microns in diameter.
Claim 14
Dependent Claim
22 words
The method of claim 1 wherein the gel particle is from between about 37 microns and about 75 microns in diameter. MS-1455-CIP
Claim 15
Dependent Claim
87 words
A substantially stable immobilized hapten reagent for use in the specific binding assay determination of a hapten or binding analog thereof in a liquid test sample, the immobilized hapten reagent comprising a gel particle which is substantially swellable in water and a plurality of hapten moieties bound thereto, said gel particle comprising a plurality of external and internal functional groups wherein substantially all of said bound hapten moieties are covalently linked to said external functional groups by a linking group which is substantially stable in aqueous solutions.
Claim 16
Dependent Claim
87 words
A substantially stable immobilized hapten reagent for use in the specific binding assay determination of a hapten or binding analog thereof in a liquid test sample, the immobilized hapten reagent comprising a gel particle which is substantially swellable in water and a plurality of hapten moieties bound thereto, said gel particle comprising a plurality of external and internal functional groups wherein substantially all of said bound hapten moieties are covalently linked to said external functional groups by a linking group which is substantially stable in aqueous solutions.
Claim 17
Dependent Claim
33 words
The immobilized hapten reagent of claim 14 wherein said gel particle is selected from the group consisting of polymers and copolymers of acrylic acid, agarose, cellulose, dextran, and their derivatives or mixtures thereof.
Claim 18
Dependent Claim
24 words
The immobilized hapten reagent of claim 14 wherein the hapten moiety is selected from the group consisting of drugs, metabolites, hormones, vitamins, and toxins.
Claim 19
Dependent Claim
24 words
The immobilized hapten reagent of claim 14 wherein the hapten moiety is selected from the group consisting of digoxin, digitoxigenin, digitoxin, digoxigenin, and 12-0-acetyldigoxigenin.
Claim 20
Dependent Claim
16 words
The immobilized hapten reagent of claim 14 wherein the hapten moiety is a glycosylated peptide sequence.
Claim 21
Dependent Claim
37 words
The immobilized hapten reagent of claim 14 wherein less than from about 1×10 -12 moles hapten/gram gel particle dissociates from the gel particle of the immobilized hapten reagent upon standing in an aqueous solution for one week.
Claim 22
Dependent Claim
37 words
The immobilized hapten reagent of claim 19 wherein less than from about 1×10 -13 moles hapten/gram gel particle dissociates from the gel particle of the immobilized hapten reagent upon standing in an aqueous solution for one week.
Claim 23
Dependent Claim
17 words
The method of claim 14 wherein the gel particle is less than about 1,000 microns in diameter.
Claim 24
Dependent Claim
21 words
The method of claim 14 wherein the gel particle is from between about 0.001 microns and about 850 microns in diameter.
Claim 25
Dependent Claim
21 words
The method of claim 14 wherein the gel particle, is from between about 37 microns and about 75 microns in diameter.
Claim 26
Dependent Claim
171 words
In an immunometric assay method for determining a hapten or analog thereof in a liquid test sample, wherein the test sample is contacted with a labeled antibody reagent capable of binding with said hapten or analog thereof and with an immobilized form of the hapten capable of binding with said antibody reagent, and wherein the amount of labeled antibody reagent which becomes bound to said hapten or analog thereof in the sample compared to that which becomes bound to the immobilized reagent is determined and related to the presence of the hapten or analog thereof in the test sample, the improvement which comprises employing as the immobilized reagent a substantially stable immobilized hapten reagent comprising a gel particle which is substantially swellable in water and a plurality of hapten moieties bound thereto, said gel particle comprising a plurality of external and internal functional groups wherein substantially all of said bound hapten moieties are covalently linked to said external functional groups by a linking group which is substantially stable in aqueous solutions.
Claim 27
Dependent Claim
171 words
In an immunometric assay method for determining a hapten or analog thereof in a liquid test sample, wherein the test sample is contacted with a labeled antibody reagent capable of binding with said hapten or analog thereof and with an immobilized form of the hapten capable of binding with said antibody reagent, and wherein the amount of labeled antibody reagent which becomes bound to said hapten or analog thereof in the sample compared to that which becomes bound to the immobilized reagent is determined and related to the presence of the hapten or analog thereof in the test sample, the improvement which comprises employing as the immobilized reagent a substantially stable immobilized hapten reagent comprising a gel particle which is substantially swellable in water and a plurality of hapten moieties bound thereto, said gel particle comprising a plurality of external and internal functional groups wherein substantially all of said bound hapten moieties are covalently linked to said external functional groups by a linking group which is substantially stable in aqueous solutions.
Claim 28
Dependent Claim
31 words
The method of claim 24 wherein said gel particle is selected from the group consisting of polymers and copolymers of acrylic acid, agarose, cellulose, dextran, and their derivatives or mixtures thereof.
Claim 29
Dependent Claim
22 words
The method of claim 24 wherein the hapten moiety is selected from the group consisting of drugs, metabolites, hormones, vitamins and toxins.
Claim 30
Dependent Claim
23 words
The assay method of claim 24 wherein the hapten moiety is selected from the group consisting of digoxin, digitoxigenin, digitoxin, digoxigenin, and 12-0-acetyldigoxigenin.
Claim 31
Dependent Claim
15 words
The assay method of claim 24 wherein the hapten moiety is a glycosylated peptide sequence.
Claim 32
Dependent Claim
36 words
The assay method of claim 24 wherein less than from about 1×10 -12 moles hapten/gram gel particle dissociates from the gel particle of the immobilized hapten reagent upon standing in an aqueous solution for one week.
Claim 33
Dependent Claim
36 words
The assay method of claim 29 wherein less than from about 1×10 -13 moles hapten/gram gel particle dissociates from the gel particle of the immobilized hapten reagent upon standing in an aqueous solution for one week.
Claim 34
Dependent Claim
17 words
The method of claim 24 wherein the gel particle is less than about 1,000 microns in diameter.
Claim 35
Dependent Claim
21 words
The method of claim 24 wherein the gel particle is from between about 0.001 microns and about 850 microns in diameter.
Claim 36
Dependent Claim
21 words
The method of claim 24 wherein the gel particle is from between about 37 microns and about 75 microns in diameter.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-09-02 | 1995-03-04 | 1995-09-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-09-02 | 1999-03-04 | 1999-09-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-09-02 | 2003-03-04 | 2003-09-03 | $7700.00 | Window Closed / Lapsed |