Patent US4148869A successfully refreshed from Google Patents.
US4148869A
Expired / Lapsed
US
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Immunological reagent and method of using same
Assignee: Bayer Corp
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Filing: 1975-09-19
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Granted: 1979-04-10
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Expires: 1995-09-19
Abstract
Abstract Immunological assaying methods which require insolubilization of an antigen-antibody complex as an integral step thereof are improved by adding to the test medium in which insolubilization is desired an aqueous reagent solution containing from about 3 to 6% by weight of a mixture of polyethylene glycol and a nonionic surfactant, and which solution has a calculated HLB (Hydrophilic-Lipophilic-Balance) of about 0.7 to 1.7. This solution increases the insolubilization of the antigen-antibody complex with reduced incubation times, and produces an assay procedure of greater test range and sensitivity. A preferred embodiment involves usage in a nephelometric analysis.
Inventors
- 👤 Carlton D. Deaton
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:02
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/909; Y10S530/00; Y10S530/806
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
65 words
A reagent for clinical nephelometric analysis comprising an aqueous solution of an antiserum to the component to be assayed and a mixture of about 20% to about 40% by weight polyethylene glycol having a molecular weight of about 200 to about 10,000 and about 80% to about 60% by weight of a block copolymer of ethylene oxide and polyoxypropylene containing at least 50% ethylene oxide.
Claim 2
Dependent Claim
65 words
A reagent for clinical nephelometric analysis comprising an aqueous solution of an antiserum to the component to be assayed and a mixture of about 20% to about 40% by weight polyethylene glycol having a molecular weight of about 200 to about 10,000 and about 80% to about 60% by weight of a block copolymer of ethylene oxide and polyoxypropylene containing at least 50% ethylene oxide.
Claim 3
Dependent Claim
37 words
The reagent of claim 1 in which the concentration of said mixture is about 4% by weight, and wherein said mixture contains about 25% by weight polyethylene glycol and about 75% by weight of said block copolymer.
Claim 4
Dependent Claim
21 words
The reagent of claim 1 in which the concentration of said mixture is from about 3% to about 6% by weight.
Claim 5
Dependent Claim
18 words
The reagent of claim 3 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 6
Dependent Claim
103 words
In an immunological assaying method which involves a reaction between an antigen and an antibody to form an antigen-antibody complex, the improvement which comprises carrying out said reaction in the presence of a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 7
Dependent Claim
103 words
In an immunological assaying method which involves a reaction between an antigen and an antibody to form an antigen-antibody complex, the improvement which comprises carrying out said reaction in the presence of a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 8
Dependent Claim
14 words
The method of claim 5 wherein said reagent is utilized in a nephelometric analysis.
Claim 9
Dependent Claim
13 words
The method of claim 5 wherein said reagent is utilized in a radioimmunoassay.
Claim 10
Dependent Claim
14 words
The method of claim 5 wherein said reagent is utilized in an enzymatic assay.
Claim 11
Dependent Claim
14 words
The method of claim 5 wherein said reagent is utilized in an electrophoretic assay.
Claim 12
Dependent Claim
18 words
The method of claim 5 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 13
Dependent Claim
103 words
In a nephelometric immunoassay of a biological constituent which involves a reaction between said constituent with its antibody, the improvement comprising incubating said constituent, prior to conducting the light scattering measurements, in a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 14
Dependent Claim
103 words
In a nephelometric immunoassay of a biological constituent which involves a reaction between said constituent with its antibody, the improvement comprising incubating said constituent, prior to conducting the light scattering measurements, in a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 15
Dependent Claim
18 words
The method of claim 11 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 16
Dependent Claim
35 words
The method of claim 11 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000 and said mixture contains about 15% to 85% polyethylene glycol and about 15% to 85% nonionic surfactant.
Claim 17
Dependent Claim
99 words
In a nephelometric immunoassay of a biological constituent, the improvement comprising incubating said constituent, prior to conducting the light scattering measurements, in a reagent comprising an aqueous solution containing an antiserum to said constituent and about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 18
Dependent Claim
99 words
In a nephelometric immunoassay of a biological constituent, the improvement comprising incubating said constituent, prior to conducting the light scattering measurements, in a reagent comprising an aqueous solution containing an antiserum to said constituent and about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 19
Dependent Claim
18 words
The method of claim 14 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 20
Dependent Claim
84 words
In a nephelometric analysis of an immunoglobulin which involves a reaction between said immunoglobulin with its antibody, the improvement comprising incubating said immunoglobulin, prior to conducting the light scattering measurements, in a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of about 20% to about 40% by weight polyethylene glycol having a molecular weight of about 200 to about 10,000 and about 80 to about 60% by weight of a block copolymer of ethylene oxide and polyoxypropylene.
Claim 21
Dependent Claim
84 words
In a nephelometric analysis of an immunoglobulin which involves a reaction between said immunoglobulin with its antibody, the improvement comprising incubating said immunoglobulin, prior to conducting the light scattering measurements, in a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of about 20% to about 40% by weight polyethylene glycol having a molecular weight of about 200 to about 10,000 and about 80 to about 60% by weight of a block copolymer of ethylene oxide and polyoxypropylene.
Claim 22
Dependent Claim
18 words
The method of claim 16 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 23
Dependent Claim
125 words
In an immunological assaying method which involves a reaction between an antigen and an antibody to form an antigen-antibody complex, the improvement which comprises carrying out said reaction in the presence of a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant selected from the group consisting of: (a) a block copolymer of ethylene oxide and polyoxypropylene, (b) straight chain primary aliphatic oxyalkylated alcohols, and (c) glycerol monostearate, wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 24
Dependent Claim
125 words
In an immunological assaying method which involves a reaction between an antigen and an antibody to form an antigen-antibody complex, the improvement which comprises carrying out said reaction in the presence of a reagent comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant selected from the group consisting of: (a) a block copolymer of ethylene oxide and polyoxypropylene, (b) straight chain primary aliphatic oxyalkylated alcohols, and (c) glycerol monostearate, wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 25
Dependent Claim
18 words
The method of claim 18 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 26
Dependent Claim
103 words
A reagent for use in an immunological assay comprising an aqueous solution containing a mixture of (1) polyethylene glycol having a molecular weight of about 4,000 to about 6,000, (2) a block copolymer of ethylene and polyoxypropylene and (3) an antiserum to the component to be assayed, wherein the combined amount of polyethylene glycol and block copolymer present in the solution is about 3 to 6% by weight and comprises about 20% to about 40% by weight polyethylene glycol and about 80% to about 60% by weight block copolymer, and wherein the calculated HLB value of the solution is about 0.7 to 1.7.
Claim 27
Dependent Claim
103 words
A reagent for use in an immunological assay comprising an aqueous solution containing a mixture of (1) polyethylene glycol having a molecular weight of about 4,000 to about 6,000, (2) a block copolymer of ethylene and polyoxypropylene and (3) an antiserum to the component to be assayed, wherein the combined amount of polyethylene glycol and block copolymer present in the solution is about 3 to 6% by weight and comprises about 20% to about 40% by weight polyethylene glycol and about 80% to about 60% by weight block copolymer, and wherein the calculated HLB value of the solution is about 0.7 to 1.7.
Claim 28
Dependent Claim
27 words
The reagent of claim 20 wherein the antiserum is an antiserum to a member selected from the group consisting of IgG, IgM, IgA, complement C3 and transferrin.
Claim 29
Dependent Claim
101 words
A reagent for use in an immunoassay comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol, said mixture containing about 10 to 90% by weight polyethylene glycol and about 10 to 90% by weight nonionic surfactant, and wherein said solution has a calculated HLB value of about 0.7 to 1.7 provided, however, that when the nonionic surfactant is a block copolymer of ethylene oxide and polyoxypropylene the block copolymer contains at least 50% ethylene oxide.
Claim 30
Dependent Claim
101 words
A reagent for use in an immunoassay comprising an aqueous solution containing about 3 to 6% by weight of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol, said mixture containing about 10 to 90% by weight polyethylene glycol and about 10 to 90% by weight nonionic surfactant, and wherein said solution has a calculated HLB value of about 0.7 to 1.7 provided, however, that when the nonionic surfactant is a block copolymer of ethylene oxide and polyoxypropylene the block copolymer contains at least 50% ethylene oxide.
Claim 31
Dependent Claim
18 words
The reagent of claim 22 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 32
Dependent Claim
118 words
In a nephelometric immunoassay of a biological constituent wherein said biological constituent undergoes an immunoreaction, the improvement comprising diluting an aqueous solution containing polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol to produce a diluted solution containing about 3 to 6% by weight of a mixture of polyethylene glycol and said nonionic surfactant having a calculated HLB value of about 0.7 to 1.7, said mixture containing about 10-90% by weight polyethylene glycol and about 10-90% by weight nonionic surfactant, and contacting said biological constituent with said diluted solution either prior to or at the time of said immunoreaction and prior to conducting the light scattering measurements.
Claim 33
Dependent Claim
118 words
In a nephelometric immunoassay of a biological constituent wherein said biological constituent undergoes an immunoreaction, the improvement comprising diluting an aqueous solution containing polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol to produce a diluted solution containing about 3 to 6% by weight of a mixture of polyethylene glycol and said nonionic surfactant having a calculated HLB value of about 0.7 to 1.7, said mixture containing about 10-90% by weight polyethylene glycol and about 10-90% by weight nonionic surfactant, and contacting said biological constituent with said diluted solution either prior to or at the time of said immunoreaction and prior to conducting the light scattering measurements.
Claim 34
Dependent Claim
18 words
The immunoassay of claim 24 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 35
Dependent Claim
42 words
The method of claim 24 wherein the biological constituent is selected from the group consisting of IgG, IgA, IgM, transferrin, complement C3, haptoglobin, alpha 1 -antitrypsin, albumin, alpha 2 -macroglobulin, alpha 1 -acid glycoprotein, T-3, T-4, triglycerides, human chorionic ganodotropins and lipoproteins.
Claim 36
Dependent Claim
35 words
The immunoassay of claim 24 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000 and said mixture contains about 15% to 85% polyethylene glycol and about 15% to 85% nonionic surfactant.
Claim 37
Dependent Claim
18 words
The immunoassay of claim 27 wherein the nonionic surfactant is a block copolymer of ethylene oxide and polyoxypropylene.
Claim 38
Dependent Claim
115 words
In a nephelometric immunoassay of a biological constituent, the improvement comprising diluting with an antiserum for said biological constituent an aqueous solution containing polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol to produce a diluted solution containing about 3 to 6% by weight of a mixture of polyethylene glycol and said nonionic surfactant wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7, and contacting said biological constituent with said diluted solution prior to conducting the light scattering measurements.
Claim 39
Dependent Claim
115 words
In a nephelometric immunoassay of a biological constituent, the improvement comprising diluting with an antiserum for said biological constituent an aqueous solution containing polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol to produce a diluted solution containing about 3 to 6% by weight of a mixture of polyethylene glycol and said nonionic surfactant wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7, and contacting said biological constituent with said diluted solution prior to conducting the light scattering measurements.
Claim 40
Dependent Claim
42 words
The method of claim 29 wherein the biological constituent is selected from the group consisting of IgG, IgA, IgM, transferrin, complement C3, haptoglobin, alpha 1 -antitrypsin, albumin, alpha 2 -macroglobulin, alpha 1 -acid glycoprotein, T-3, T-4, triglycerides, human chorionic ganadotropins and lipoproteins.
Claim 41
Dependent Claim
18 words
The immunoassay of claim 29 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 42
Dependent Claim
12 words
The immunoassay of claim 41 wherein the antiserum is a prediluted antiserum.
Claim 43
Dependent Claim
18 words
The immunoassay of claim 32 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 44
Dependent Claim
112 words
In an immunoassay which produces an antigen-antibody complex, the improvement comprising diluting with an antiserum for said antigen or antibody an aqueous solution containing polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol to produce a diluted solution containing about 3% to 6% by weight of a mixture of polyethylene glycol and said nonionic surfactant wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7, and producing the antigen-antibody complex in the presence of said diluted solution.
Claim 45
Dependent Claim
112 words
In an immunoassay which produces an antigen-antibody complex, the improvement comprising diluting with an antiserum for said antigen or antibody an aqueous solution containing polyethylene glycol having a molecular weight of about 200 to about 10,000 and a nonionic surfactant other than polyethylene glycol to produce a diluted solution containing about 3% to 6% by weight of a mixture of polyethylene glycol and said nonionic surfactant wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight nonionic surfactant and said solution has a calculated HLB value of about 0.7 to 1.7, and producing the antigen-antibody complex in the presence of said diluted solution.
Claim 46
Dependent Claim
42 words
The method of claim 34 wherein the biological component is selected from the group consisting of IgG, IgA, IgM, transferrin, complement C3, haptoglobin, alpha 1 -antitrypsin, albumin, alpha 2 -macroglobulin, alpha 1 -acid glycoprotein, T-3, T-4, triglycerides, human chorionic ganadotropins and lipoproteins.
Claim 47
Dependent Claim
51 words
The method of claim 34 wherein the nonionic surfactant is a block copolymer of ethylene oxide and polyoxypropylene, the calculated HLB value of the diluted solution is about 0.7 to 1.3, and the biological component is selected from the group consisting of IgG, IgA, IgM, complement C3, transferrin, T3 and T4.
Claim 48
Dependent Claim
18 words
The immunoassay of claim 34 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 49
Dependent Claim
12 words
The immunoassay of claim 34 wherein the antiserum is a prediluted antiserum.
Claim 50
Dependent Claim
18 words
The immunoassay of claim 38 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
Claim 51
Dependent Claim
99 words
In an immunological assaying method which involves a reaction between an antigen and an antibody to form an antigen-antibody complex, the improvement which comprises carrying out said reaction in the presence of a reagent comprising an aqueous solution containing about 3% to 6% of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and an alkyl aryl sulfonate wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight alkyl aryl sulfonate and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 52
Dependent Claim
99 words
In an immunological assaying method which involves a reaction between an antigen and an antibody to form an antigen-antibody complex, the improvement which comprises carrying out said reaction in the presence of a reagent comprising an aqueous solution containing about 3% to 6% of a mixture of polyethylene glycol having a molecular weight of about 200 to about 10,000 and an alkyl aryl sulfonate wherein said mixture contains about 10% to 90% by weight polyethylene glycol and about 10% to 90% by weight alkyl aryl sulfonate and said solution has a calculated HLB value of about 0.7 to 1.7.
Claim 53
Dependent Claim
14 words
The method of claim 40 wherein the immunological assaying method is a nephelometric analysis.
Claim 54
Dependent Claim
18 words
The method of claim 40 wherein the polyethylene glycol has a molecular weight of about 4,000 to 6,000.
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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) | 1982-04-09 | 1982-10-09 | 1983-04-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1986-04-09 | 1986-10-09 | 1987-04-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1990-04-09 | 1990-10-09 | 1991-04-10 | $7700.00 | Window Closed / Lapsed |