US4226935A
Expired / Lapsed
US
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Enzymatic diagnostic composition
Assignee: WR Grace and Co Conn
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Filing: 1978-08-07
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Granted: 1980-10-07
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Expires: 1998-08-07
Abstract
Abstract Disclosed herein is a method for binding enzymes comprising reacting an amino acid and/or a protein with an excess of a urethane prepolymer, curing the resulting product and coupling an enzyme thereto by use of a carbodiimide. The product of the process is also described as well as the use thereof in analyzing blood fluids. The invention is particularly adapted to binding hexokinase and glucose-6-phosphate dehydrogenase to determine creatine phosphokinase (CPK) in human blood serum.
Inventors
- 👤 Murray C. Fusee
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 04:35
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/48; C08; C08G; C08G18/00; C08G18/06; C08G18/08; C08G18/10; C08G18/28; C08G18/40; C08G18/48; C08G18/4833; C08G18/64; C08G18/6415; C08G18/6446; C12N; C12N11/00; C12N11/02; C12N11/08; C12N11/089; C12N11/093; C12N11/098; C12N11/18; C12Q1/26; C12Q1/32
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
71 words
A diagnostic composition comprising a mixture of hexokinase and glucose-6-phosphate dehydrogenase bound in biologically-active form to a carboxylated poly(urea-urethane) polymer matrix having oxyalkylene backbone segments wherein at least 60 mole percent of the oxyalkylene units are oxyethylene said carboxylated polymer prepared by reacting an excess of a polyoxyalkylene urethane prepolymer with a monomeric amino acid, and wherein said enzymes are bonded to at least one exterior surface of said polymer matrix.
Claim 2
Dependent Claim
71 words
A diagnostic composition comprising a mixture of hexokinase and glucose-6-phosphate dehydrogenase bound in biologically-active form to a carboxylated poly(urea-urethane) polymer matrix having oxyalkylene backbone segments wherein at least 60 mole percent of the oxyalkylene units are oxyethylene said carboxylated polymer prepared by reacting an excess of a polyoxyalkylene urethane prepolymer with a monomeric amino acid, and wherein said enzymes are bonded to at least one exterior surface of said polymer matrix.
Claim 3
Dependent Claim
35 words
A composition as in claim 1 wherein, based on the total weight of the composition, from about 0.001 to about 1.0 percent is hexokinase and from about 0.001 to about 1.0 percent is glucose-6-phosphate dehydrogenase.
Claim 4
Dependent Claim
16 words
A composition as in claim 1 wherein the matrix is in the form of a foam.
Claim 5
Dependent Claim
16 words
A composition as in claim 1 wherein the matrix is in the form of a film.
Claim 6
Dependent Claim
26 words
A composition as in claim 1 wherein the matrix is in the form of a self-supporting tubular film having enzymes bound on the interior surface thereof.
Claim 7
Dependent Claim
13 words
A composition as in claim 1 wherein the backbone components are polyoxyethylene/polyoxypropylene copolymers.
Claim 8
Independent Claim
89 words
A method for analyzing human body fluids comprising contacting a first aqueous solution of said body fluids with a mixture of hexokinase and glucose-6-phosphate dehydrogenase bound in biologically-active form to a carboxylated polyurethane polymer matrix, allowing said immobilized enzymes to catalyze reaction between the components of the first aqueous solution to form a second aqueous solution and determining the amount of at least one component of said second aqueous solution, said polymer comprising the reaction product of an excess of a polyoxyalkylene urethane prepolymer and a monomeric amino acid.
Claim 9
Independent Claim
89 words
A method for analyzing human body fluids comprising contacting a first aqueous solution of said body fluids with a mixture of hexokinase and glucose-6-phosphate dehydrogenase bound in biologically-active form to a carboxylated polyurethane polymer matrix, allowing said immobilized enzymes to catalyze reaction between the components of the first aqueous solution to form a second aqueous solution and determining the amount of at least one component of said second aqueous solution, said polymer comprising the reaction product of an excess of a polyoxyalkylene urethane prepolymer and a monomeric amino acid.
Claim 10
Dependent Claim
19 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of adenosine triphosphate.
Claim 11
Dependent Claim
19 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of adenosine diphosphate.
Claim 12
Dependent Claim
19 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of creatine phosphate.
Claim 13
Dependent Claim
18 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of glucose.
Claim 14
Dependent Claim
18 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of glucose-6-phosphate.
Claim 15
Dependent Claim
20 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of nicotinamide adenine dinucleotide.
Claim 16
Dependent Claim
21 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of nicotinamide adenine dinucleotide phosphate.
Claim 17
Dependent Claim
18 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of glucose-1-phosphate.
Claim 18
Dependent Claim
18 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of fructose-6-phosphate.
Claim 19
Dependent Claim
18 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of myokinase.
Claim 20
Dependent Claim
19 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of phosphoglucose isomerase.
Claim 21
Dependent Claim
18 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of galactokinase.
Claim 22
Dependent Claim
19 words
A method as in claim 7 wherein the body fluid is analyzed to determine the presence of creatine phosphokinase.
Claim 23
Dependent Claim
53 words
A method as in claim 20 wherein the first aqueous solution comprises the body fluid to be tested, creatine phosphate, glucose, adenosine diphosphate, magnesium ions, and NAD or NAD phosphate, and the second aqueous solution comprises NADH or NADPH in an amount proportional to the creatine phosphokinase level in the body fluid tested.
Claim 24
Dependent Claim
41 words
A method as in claim 20 wherein the polymer matrix comprises, based on the total weight of said matrix and enzymes bound therein, from about 0.001 to about 1% of hexokinase and from about 0.001 to about 1% of glucose-6-phosphate dehydrogenase.
Claim 25
Dependent Claim
17 words
A method as in claim 20 wherein the polymer matrix is in the form of a foam.
Claim 26
Dependent Claim
13 words
A method as in claim 20 wherein the polymer matrix is a film.
Claim 27
Dependent Claim
15 words
A method as in claim 20 wherein the polymer matrix is a tubular self-supporting film.
Claim 28
Dependent Claim
15 words
A method as in claim 20 wherein the NADH or NADPH concentration is determined photometrically.
Claim 29
Independent Claim
38 words
A method for binding enzymes comprising reacting a monomeric amino acid with an excess of a polyoxyalkylene urethane prepolymer to form a carboxylated prepolymer, curing the carboxylated prepolymer and coupling an enzyme thereto by use of a carbodiimide.
Claim 30
Independent Claim
38 words
A method for binding enzymes comprising reacting a monomeric amino acid with an excess of a polyoxyalkylene urethane prepolymer to form a carboxylated prepolymer, curing the carboxylated prepolymer and coupling an enzyme thereto by use of a carbodiimide.
Claim 31
Dependent Claim
18 words
A method as in claim 27 including the additional step of shaping the carboxylated prepolymer prior to curing.
Claim 32
Dependent Claim
16 words
A method as in claim 27 wherein the carboxylated prepolymer is cured to form a foam.
Claim 33
Dependent Claim
16 words
A method as in claim 27 wherein the carboxylated prepolymer is cured to form a film.
Claim 34
Dependent Claim
12 words
A method as in claim 27 wherein the enzyme employed is hexokinase.
Claim 35
Dependent Claim
13 words
A method as in claim 27 wherein the enzyme employed is glucose-6-phosphate dehydrogenase.
Claim 36
Dependent Claim
16 words
A method as in claim 27 wherein a mixture of hexokinase and glucose-6-phosphate dehydrogenase is employed.
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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) | 1983-10-07 | 1984-04-07 | 1984-10-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-10-07 | 1988-04-07 | 1988-10-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-10-07 | 1992-04-07 | 1992-10-07 | $7700.00 | Window Closed / Lapsed |