US4921809A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Polymer coated solid matrices and use in immunoassays
Assignee: Bostik Inc
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Filing: 1987-09-29
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Granted: 1990-05-01
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Expires: 2007-09-29
Abstract
Abstract Certain polyethyleneimines are used to coat solid carriers used in immunoassays to improve the sensitivity of the assay over what is obtained without the coating. Use of the invention in enzyme immunoassays yields sensitivities equal to that of radioimmunoassays. A post-coating is also used after the solid carrier has had a layer of capture molecule applied and before the assay is performed, to improve even further the sensitivity, reliability and dynamic range of the assay results.
Inventors
- 👤 Robert Schiff
- 👤 Gary Lehnus
- 👤 Charles J. Wasserman
- 👤 Henrietta M. Garlock
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 17:06
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54393; G01N33/544; G01N33/545; G01N33/68; G01N33/6854
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
70 words
In an immunoassay solid support carrying a capture molecule directly or through an interposing layer of a material facilitating covalent or adsorptive bonding of the capture molecule to the support, the improvement which comprises a layer of a polyethyleneimine on said support intermediate said support and said capture molecule and wherein there is a blocking agent present on the polyethyleneimine capture antibody composite comprising a mixture of ovalbumin and dithiothreitol.
Claim 2
Dependent Claim
70 words
In an immunoassay solid support carrying a capture molecule directly or through an interposing layer of a material facilitating covalent or adsorptive bonding of the capture molecule to the support, the improvement which comprises a layer of a polyethyleneimine on said support intermediate said support and said capture molecule and wherein there is a blocking agent present on the polyethyleneimine capture antibody composite comprising a mixture of ovalbumin and dithiothreitol.
Claim 3
Dependent Claim
16 words
The immunoassay solid support of claim 1 wherein the capture molecule is an antibody or antigen.
Claim 4
Dependent Claim
19 words
The immunoassay solid support of claim 2 wherein the polyethyleneimine has a molecular weight of from 600 to 60,000.
Claim 5
Dependent Claim
13 words
The immunoassay solid support of claim 3 wherein the solid support is polystyrene.
Claim 6
Dependent Claim
13 words
The immunoassay solid support of claim 4 wherein said interposing layer is glutaraldehyde.
Claim 7
Dependent Claim
18 words
The immunoassay solid support of claim 4 wherein the polyethyleneimine has a molecular weight of 800 to 1800.
Claim 8
Dependent Claim
26 words
The immunoassay solid support of claim 1, wherein the blocking agent additionally contains one or more of lactalbumin, lactalbumin enzymatic hydrolyzate, para-aminobenzoic acid and pyridoxyl phosphate.
Claim 9
Dependent Claim
12 words
The product of claim 7 wherein the capture antibody is for beta-HCG.
Claim 10
Dependent Claim
90 words
In a method for producing a solid support substrate useful in performing an immunoassay wherein a capture molecule is attached to said solid support directly or through an interposing layer of a material facilitating covalent or adsorptive bonding of the capture molecule to said layer, the improvement which comprises applying a layer of a polyethyleneimine onto said support intermediate said support and said capture molecule and then treating the resulting product with a mixture comprising ovalbumin and dithiothreitol sufficient in amount to provide a blocking layer of the polyethyleneimine layer.
Claim 11
Dependent Claim
90 words
In a method for producing a solid support substrate useful in performing an immunoassay wherein a capture molecule is attached to said solid support directly or through an interposing layer of a material facilitating covalent or adsorptive bonding of the capture molecule to said layer, the improvement which comprises applying a layer of a polyethyleneimine onto said support intermediate said support and said capture molecule and then treating the resulting product with a mixture comprising ovalbumin and dithiothreitol sufficient in amount to provide a blocking layer of the polyethyleneimine layer.
Claim 12
Dependent Claim
14 words
The method of claim 9 wherein the capture molecule is an antibody or antigen.
Claim 13
Dependent Claim
17 words
The method of claim 10 wherein the polyethyleneimine has a molecular weight of from 600 to 60,000.
Claim 14
Dependent Claim
11 words
The method of claim 11 wherein the solid support is polystyrene.
Claim 15
Dependent Claim
11 words
The method of claim 12 wherein said interposing layer is glutaraldehyde.
Claim 16
Dependent Claim
11 words
The method of claim 13 wherein the support layer is polystyrene.
Claim 17
Dependent Claim
12 words
The method of claim 13 wherein the capture antibody is for beta-HCG.
Claim 18
Dependent Claim
27 words
The method of claim 13 wherein the blocking mixture further comprises one or more of the following: para-aminobenzoic acid, lactalbumin, enzymatic hydrolysate of lactalbumin, and pyridoxyl phosphate.
Claim 19
Dependent Claim
32 words
The method of claim 16 wherein the blocking mixture comprises, by weight, 45-80% ovalbumin, 0.7-1.5% dithiothreitol, 16-50% lactalbumin or lactalbumin enzymatic hydrolysate, 1.3-2.2% para-aminobenzoic acid, and 15-18% pyridoxyl phosphate in aqueous solution.
Claim 20
Dependent Claim
145 words
In a method for conducting an immunoassay wherein a solid support carrier having a capture antibody attached thereto either directly or through an interposing layer of a material facilitating covalent or adsorptive bonding of the capture antibody to the layer, and said antibody is contacted with a liquid in which the presence and/or concentration of an antigen is to be determined, and said contacted antibody is contacted with a labeled form of a binding partner of said antigen and the extent of attachment of such labeled form is measured to measure the presence and/ or concentration of said antigen, the improvement which comprises applying a layer of a polyethyleneimine to said support carrier intermediate said support and said capture antibody and then treating the resulting product with a mixture comprising ovalbumin and dithiothreitol sufficient in amount to provide a blocking layer on the polyethyleneimine layer.
Claim 21
Dependent Claim
145 words
In a method for conducting an immunoassay wherein a solid support carrier having a capture antibody attached thereto either directly or through an interposing layer of a material facilitating covalent or adsorptive bonding of the capture antibody to the layer, and said antibody is contacted with a liquid in which the presence and/or concentration of an antigen is to be determined, and said contacted antibody is contacted with a labeled form of a binding partner of said antigen and the extent of attachment of such labeled form is measured to measure the presence and/ or concentration of said antigen, the improvement which comprises applying a layer of a polyethyleneimine to said support carrier intermediate said support and said capture antibody and then treating the resulting product with a mixture comprising ovalbumin and dithiothreitol sufficient in amount to provide a blocking layer on the polyethyleneimine layer.
Claim 22
Dependent Claim
17 words
The method of claim 18 wherein the polyethyleneimine has a molecular weight of from 600 to 60,000.
Claim 23
Dependent Claim
11 words
The method of claim 19 wherein the solid support is polystyrene.
Claim 24
Dependent Claim
26 words
The method of claim 20 wherein a layer of glutaraldehyde is applied to said layer of polyethyleneimine to facilitate covalent bonding of the capture antibody thereto.
Claim 25
Dependent Claim
11 words
The method of claim 21 wherein the support layer is polystyrene.
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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) | 1993-04-30 | 1993-10-30 | 1994-05-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-04-30 | 1997-10-30 | 1998-05-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-04-30 | 2001-10-30 | 2002-05-01 | $7700.00 | Window Closed / Lapsed |