US4414326A
Expired / Lapsed
US
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Diagnostic agents
Assignee: Individual
•
Filing: 1982-02-04
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Granted: 1983-11-08
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Expires: 2002-02-04
Abstract
Abstract Diagnostic agents are described for determining high density lipoprotein cholesterol in body fuids such as serum or plasma. One is an enzymatic reagent for quantitative analysis for cholesterol composed of cholesterol oxidase, cholesterol esterase, peroxidase, 4-amino-antipyrine, a phenol or phenol derivative, a surfactant, such as sodium cholate and a polyglycol of molecular weight from about 190 to 1000, buffered to pH 5.5 to 7.8 in water. The other is a single, stable precipitating agent for lipoproteins in body fluids, comprising a bivalent metal salt of a monobasic acid, such as magnesium chloride and a water-soluble polyanion, such as sodium phosphotungstate in water at pH 5 to 8.
Inventors
- 👤 Jack M. Goldberg
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 04:24
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/60; Y; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
133 words
A stable, aqueous enzymatic reagent for interaction in the presence of cholesterol to provide a measurable chromophore, said reagent capable of remaining stable for a relatively long period of time comprising: effective amounts of cholesterol oxidase, cholesterol esterase derived from an animal source, peroxidase, 4-aminoantipyrine, an agent capable of forming a chromophore, a bile salt, a water-soluble polyglycol having an approximate weight average molecular weight in the range of 190-1000 selected from the group consisting of polyethylene glycol and polyethoxy glycol, in an amount of 0.1-1.0 grams per liter of enzymatic reagent and a stabilizer comprising a water-soluble polyglycol having an approximate weight average molecular weight of 6000 or higher in an amount sufficient to maintain solubility of free cholesterol, said enzymatic reagent having a pH in the range of 5.5 to 7.8.
Claim 2
Dependent Claim
133 words
A stable, aqueous enzymatic reagent for interaction in the presence of cholesterol to provide a measurable chromophore, said reagent capable of remaining stable for a relatively long period of time comprising: effective amounts of cholesterol oxidase, cholesterol esterase derived from an animal source, peroxidase, 4-aminoantipyrine, an agent capable of forming a chromophore, a bile salt, a water-soluble polyglycol having an approximate weight average molecular weight in the range of 190-1000 selected from the group consisting of polyethylene glycol and polyethoxy glycol, in an amount of 0.1-1.0 grams per liter of enzymatic reagent and a stabilizer comprising a water-soluble polyglycol having an approximate weight average molecular weight of 6000 or higher in an amount sufficient to maintain solubility of free cholesterol, said enzymatic reagent having a pH in the range of 5.5 to 7.8.
Claim 3
Dependent Claim
25 words
An enzymatic regent as defined in claim 1 wherein said polyglycol having an approximate weight average molecular weight of 6000 or higher is polyethylene glycol.
Claim 4
Dependent Claim
23 words
An enzymatic reagent as defined in claim 1 wherein said agent capable of forming a chromophore comprises a phenol or a phenol derivative.
Claim 5
Dependent Claim
19 words
An enzymatic reagent as defined in claim 1 wherein said chromophore forming agent comprises a phenol or phenol derivative.
Claim 6
Dependent Claim
140 words
A two component enzymatic reagent comprising a first, solid, enzymatic composition and a second, separated, liquid activator therefor, both the solid enzymatic composition and the liquid activator being substantially stable for a period of time of 6 months or more when separated, comprising: a first, solid enzymatic composition comprising enzyme effective amounts of cholesterol oxidase, cholesterol esterase derived from an animal source, peroxidase, 4-aminoantipyrine and a water-soluble polyglycol having an approximate weight average molecular weight of 6000 or higher in an amount sufficient to maintain solubility of free cholesterol, said composition having a pH in the range of 5.5 to 7.8; and a second, liquid activator comprising a bile salt and a water soluble polyglycol having a weight average molecular weight in the approximate range of 190 to 1000 selected from the group consisting of polyethylene glycol and polyethoxy glycol.
Claim 7
Dependent Claim
140 words
A two component enzymatic reagent comprising a first, solid, enzymatic composition and a second, separated, liquid activator therefor, both the solid enzymatic composition and the liquid activator being substantially stable for a period of time of 6 months or more when separated, comprising: a first, solid enzymatic composition comprising enzyme effective amounts of cholesterol oxidase, cholesterol esterase derived from an animal source, peroxidase, 4-aminoantipyrine and a water-soluble polyglycol having an approximate weight average molecular weight of 6000 or higher in an amount sufficient to maintain solubility of free cholesterol, said composition having a pH in the range of 5.5 to 7.8; and a second, liquid activator comprising a bile salt and a water soluble polyglycol having a weight average molecular weight in the approximate range of 190 to 1000 selected from the group consisting of polyethylene glycol and polyethoxy glycol.
Claim 8
Dependent Claim
25 words
An enzymatic regent as defined in claim 5 wherein said polyglycol having an approximate weight average molecular weight of 6000 or higher is polyethylene glycol.
Claim 9
Independent Claim
137 words
A method of manufacturing an enzymatic reagent having increased stability when reconstituted with water comprising admixing an aqueous solution of cholesterol oxidase, choletserol esterase derived from an animal source, peroxidase, 4-aminoantipyrine, a bile salt, a water-soluble polyglycol having an approximate weight average molecular weight of 6000 or higher in an amount sufficient to maintain solubility of free cholesterol, and a lower molecular weight water-soluble polyglycol having a weight average molecular weight in the range of about 190 to 1000 selected from the group consisting of polyethylene glycol and polyethoxy glycol, said enzymatic reagent having a pH in the range of 5.5 to 7.8, said lower molecular weight polyglycol being present in said solution in an amount of 0.01%-0.1% by total weight of said solution; and drying said aqueous solution to provide a substantially completely solid enzymatic composition.
Claim 10
Independent Claim
137 words
A method of manufacturing an enzymatic reagent having increased stability when reconstituted with water comprising admixing an aqueous solution of cholesterol oxidase, choletserol esterase derived from an animal source, peroxidase, 4-aminoantipyrine, a bile salt, a water-soluble polyglycol having an approximate weight average molecular weight of 6000 or higher in an amount sufficient to maintain solubility of free cholesterol, and a lower molecular weight water-soluble polyglycol having a weight average molecular weight in the range of about 190 to 1000 selected from the group consisting of polyethylene glycol and polyethoxy glycol, said enzymatic reagent having a pH in the range of 5.5 to 7.8, said lower molecular weight polyglycol being present in said solution in an amount of 0.01%-0.1% by total weight of said solution; and drying said aqueous solution to provide a substantially completely solid enzymatic composition.
Claim 11
Dependent Claim
24 words
A method as defined in claim 7, wherein said polyglycol having an approximate weight average molecular weight of 6000 or higher is polyethylene glycol.
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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) | 1986-11-07 | 1987-05-09 | 1987-11-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-11-07 | 1991-05-09 | 1991-11-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-11-07 | 1995-05-09 | 1995-11-08 | $7700.00 | Window Closed / Lapsed |