US4189536A
Expired / Lapsed
US
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Reagent systems, enzymatic assays and compositions therefor
Assignee: Hycel Inc
•
Filing: 1977-11-09
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Granted: 1980-02-19
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Expires: 1997-11-09
Abstract
Abstract An improved reagent system for chemical determinations and novel compositions therefor provide improved reagent stability, particularly in a dry, storage stable phase. For determinations in which hexokinase and glucose-6-phosphate dehydrogenase are indicator enzymes, glycine and taurine are combined with bovine serum albumin to provide a bulking agent for the indicator enzymes which also acts as a stabilizer. A novel salt, the tris (hydroxymethyl)aminomethane salt of 2 mercaptosuccinic acid, acts as an activator or stabilizer in chemistries utilizing the indicator enzymes, namely determinations of creatine phosphokinase or glucose. The novel salt also acts as a stabilizer in a urea nitrogen determination. Dry reagent components are bulked in triethanolammonium terephthalate. Novel CPK, glucose and serum urea nitrogen determinations are also provided.
Inventors
- 👤 Stanley E. Green
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 14:48
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/54; C12Q1/48; C12Q1/50; C12Q1/58
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
59 words
In a solid assaying material for assaying a specimen of biological fluid in which hexokinase catalyzes the phosphorylation of glucose and in which glucose-6-phosphate is oxidized in the presence of glucose-6-phosphate dehydrogenase, the improvement comprising said enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a bulking agent comprising bovine serum albumin, glycine and taurine and further comprising te compound TMS.
Claim 2
Dependent Claim
59 words
In a solid assaying material for assaying a specimen of biological fluid in which hexokinase catalyzes the phosphorylation of glucose and in which glucose-6-phosphate is oxidized in the presence of glucose-6-phosphate dehydrogenase, the improvement comprising said enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a bulking agent comprising bovine serum albumin, glycine and taurine and further comprising te compound TMS.
Claim 3
Dependent Claim
51 words
A solid assaying material for assaying a specimen of biological fluid for creatine phosphokinase comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, nicotinimide adenine dinucleotide phosphate, glucose, TMS and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 4
Dependent Claim
51 words
A solid assaying material for assaying a specimen of biological fluid for creatine phosphokinase comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, nicotinimide adenine dinucleotide phosphate, glucose, TMS and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 5
Independent Claim
99 words
A method of assaying a specimen of biological fluid for creatine phosphokinase comprising the steps of providing as assay material in a dry powder form comprising a solid, watersoluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, nicotinimide adenine dinucleotide phosphate, glucose, TMS and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen reagent mixture, and determining the change in optical density.
Claim 6
Independent Claim
99 words
A method of assaying a specimen of biological fluid for creatine phosphokinase comprising the steps of providing as assay material in a dry powder form comprising a solid, watersoluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, nicotinimide adenine dinucleotide phosphate, glucose, TMS and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen reagent mixture, and determining the change in optical density.
Claim 7
Dependent Claim
51 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, nicotinimide adenine dinucleotide, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising TMS.
Claim 8
Dependent Claim
51 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, nicotinimide adenine dinucleotide, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising TMS.
Claim 9
Independent Claim
106 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, nicotinimide adenine dinucleotide, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising TMS, and further comprising the steps of providing said material in dry powder form, dissolving said material in water to provide an aqueous reagent, mixing said specimen with said aqueous reagent and obtaining a reacted specimen reagent mixture, and measuring change in optical density of said reacted reagent-specimen mixture at a preselected wavelength.
Claim 10
Independent Claim
106 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, nicotinimide adenine dinucleotide, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising TMS, and further comprising the steps of providing said material in dry powder form, dissolving said material in water to provide an aqueous reagent, mixing said specimen with said aqueous reagent and obtaining a reacted specimen reagent mixture, and measuring change in optical density of said reacted reagent-specimen mixture at a preselected wavelength.
Claim 11
Dependent Claim
25 words
In a bulking agent for hexokinase comprising bovine serum albumin, the improvement wherein said bulking agent further comprises a mixture consisting of glycine and taurine.
Claim 12
Dependent Claim
25 words
In a bulking agent for hexokinase comprising bovine serum albumin, the improvement wherein said bulking agent further comprises a mixture consisting of glycine and taurine.
Claim 13
Dependent Claim
26 words
In a bulking agent for glucose-6-phosphate dehydrogenase comprising bovine serum albumin, the improvement wherein said bulking agent further comprises a mixture consisting of glycine and taurine.
Claim 14
Dependent Claim
26 words
In a bulking agent for glucose-6-phosphate dehydrogenase comprising bovine serum albumin, the improvement wherein said bulking agent further comprises a mixture consisting of glycine and taurine.
Claim 15
Dependent Claim
28 words
In a bulking agent for hexokinase and glucose-6-phosphate dehydrogenase comprising bovine serum albumin, the improvement wherein said bluking agent further comprises a mixture consisting of glycine and taurine.
Claim 16
Dependent Claim
28 words
In a bulking agent for hexokinase and glucose-6-phosphate dehydrogenase comprising bovine serum albumin, the improvement wherein said bluking agent further comprises a mixture consisting of glycine and taurine.
Claim 17
Dependent Claim
25 words
In a bulking agent for diaphorase comprising bovine serum albumin, the improvement wherein said bulking agent further comprises a mixture consisting of glycine and taurine.
Claim 18
Dependent Claim
25 words
In a bulking agent for diaphorase comprising bovine serum albumin, the improvement wherein said bulking agent further comprises a mixture consisting of glycine and taurine.
Claim 19
Dependent Claim
54 words
A solid assaying material for assaying a specimen of biological fluid for creatine phosphokinase comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, a nicotinamide adenine dinucleotide coenzyme, glucose, a sulfhydryl activator and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 20
Dependent Claim
54 words
A solid assaying material for assaying a specimen of biological fluid for creatine phosphokinase comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, a nicotinamide adenine dinucleotide coenzyme, glucose, a sulfhydryl activator and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 21
Independent Claim
110 words
A method of assaying a specimen of biological fluid for creatine phosphokinase comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, nicotinimide adenine dinucleotide phosphate, glucose, a sulfhydryl activator and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum ablumin, glycine and taurine, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 22
Independent Claim
110 words
A method of assaying a specimen of biological fluid for creatine phosphokinase comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, nicotinimide adenine dinucleotide phosphate, glucose, a sulfhydryl activator and hexokinase and glycerol-6-phosphate dehydrogenase bulked in a mixture of bovine serum ablumin, glycine and taurine, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 23
Dependent Claim
51 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, a sulfhydryl stabilizer, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 24
Dependent Claim
51 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, a sulfhydryl stabilizer, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 25
Independent Claim
96 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, a sulfhydryl stabilizer, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising the steps of dissolving said material to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted reagent-specimen mixture and measuring change in optical density of said reacted reagent-specimen mixture at a preselected wavelength.
Claim 26
Independent Claim
96 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, a sulfhydryl stabilizer, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine, and further comprising the steps of dissolving said material to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted reagent-specimen mixture and measuring change in optical density of said reacted reagent-specimen mixture at a preselected wavelength.
Claim 27
Dependent Claim
40 words
A solid assaying material for assaying a specimen of biological fluid for creatine phosphokinase comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, a nicotinimide adenine dinucleotide coenzyme, glucose, TMS, hexokinase and glycerol-6-phosphate dehydrogenase.
Claim 28
Dependent Claim
40 words
A solid assaying material for assaying a specimen of biological fluid for creatine phosphokinase comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, a nicotinimide adenine dinucleotide coenzyme, glucose, TMS, hexokinase and glycerol-6-phosphate dehydrogenase.
Claim 29
Independent Claim
97 words
A method of assaying a specimen of biological fluid for creatine phosphokinase comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, a nicotinimide adenine dinocleotide coenzyme, glucose, TMS, hexokinase and glycerol-6-phosphate dehydrogenase, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 30
Independent Claim
97 words
A method of assaying a specimen of biological fluid for creatine phosphokinase comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of creatine phosphate, adenosine diphosphate, activator, a nicotinimide adenine dinocleotide coenzyme, glucose, TMS, hexokinase and glycerol-6-phosphate dehydrogenase, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 31
Dependent Claim
39 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphospahte, a nicotinimide adenine dinucleotide coenzyme, activator TMS, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase.
Claim 32
Dependent Claim
39 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphospahte, a nicotinimide adenine dinucleotide coenzyme, activator TMS, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase.
Claim 33
Independent Claim
86 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, TMS, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 34
Independent Claim
86 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, TMS, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 35
Dependent Claim
35 words
A solid assaying material for assaying a specimen of biological for urea nitrogen comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alpha-ketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, glutamate dehydrogenase and TMS.
Claim 36
Dependent Claim
35 words
A solid assaying material for assaying a specimen of biological for urea nitrogen comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alpha-ketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, glutamate dehydrogenase and TMS.
Claim 37
Independent Claim
93 words
A method of assaying a specimen of biological fluid for urea nitrogen comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alpha-ketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, glutamate dehydrogenase and TMS, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 38
Independent Claim
93 words
A method of assaying a specimen of biological fluid for urea nitrogen comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alpha-ketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, glutamate dehydrogenase and TMS, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 39
Dependent Claim
45 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixrture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in TEA-TPA to provide bulked reagent.
Claim 40
Dependent Claim
45 words
A solid assaying material for assaying a specimen of biological fluid for glucose comprising a solid, water-soluble, substantially anhydrous, storage stable mixrture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in TEA-TPA to provide bulked reagent.
Claim 41
Dependent Claim
27 words
A solid assaying material according to claim 20 wherein said bulked reagent comprises the enzymes hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin.
Claim 42
Dependent Claim
11 words
A solid assaying material according to claim 20 further comprising TMS.
Claim 43
Dependent Claim
11 words
A solid assaying material according to claim 21 further comprising TMS.
Claim 44
Independent Claim
97 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase all bulked in TEA-TPA, and further comprising the steps of providing said material in dry powder form, dissolving said material in water to provide an aqueous reagent, mixing said specimen with said aqueous reagent and obtaining a reacted specimen reagent mixture, and measuring change in optical density of said reacted reagent-specimen mixture at a preselected wavelength.
Claim 45
Independent Claim
97 words
A method of assaying a specimen for glucose comprising the step of providing an assay material comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of adenosine triphosphate, a nicotinimide adenine dinucleotide coenzyme, activator, and the enzymes hexokinase and glucose-6-phosphate dehydrogenase all bulked in TEA-TPA, and further comprising the steps of providing said material in dry powder form, dissolving said material in water to provide an aqueous reagent, mixing said specimen with said aqueous reagent and obtaining a reacted specimen reagent mixture, and measuring change in optical density of said reacted reagent-specimen mixture at a preselected wavelength.
Claim 46
Dependent Claim
32 words
A method according to claim 24 wherein the step of providing said assay material comprises providing said hexokinase and glucose-6-phosphate dehydrogenase bulked in a mixture of bovine serum albumin, glycine and taurine.
Claim 47
Dependent Claim
21 words
A method according to claim 24 wherein the step of providing said assay material further comprises the step of providing TMS.
Claim 48
Dependent Claim
21 words
A method according to claim 25 wherein the step of providing said assay material further comprises the step of providing TMS.
Claim 49
Dependent Claim
36 words
A solid assaying material for assaying a specimen for urea nitrogen comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alphaketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, and glutamate dehydrogenase, all bulked in TEA-TPA.
Claim 50
Dependent Claim
36 words
A solid assaying material for assaying a specimen for urea nitrogen comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alphaketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, and glutamate dehydrogenase, all bulked in TEA-TPA.
Claim 51
Dependent Claim
15 words
A solid assaying material according to claim 28 further comprising TMS bulked in said TEA-TPA.
Claim 52
Independent Claim
96 words
A method of assaying a specimen of biological fluid for urea nitrogen comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alpha-ketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, and glutamate dehydrogenase, all bulked in TEA-TPA, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 53
Independent Claim
96 words
A method of assaying a specimen of biological fluid for urea nitrogen comprising the step of providing an assay material provided in a dry powder form comprising a solid, water-soluble, substantially anhydrous, storage stable mixture of alpha-ketogluterate, reduced nicotinimide adenine dinucleotide, adenosine diphosphate, urease, and glutamate dehydrogenase, all bulked in TEA-TPA, and further comprising the steps of dissolving said material in water to provide an aqueous reagent, mixing said specimen in said aqueous reagent and obtaining a reacted specimen-reagent mixture, and determining the change in optical density of the reacted specimen-reagent mixture at a preselected wavelength.
Claim 54
Dependent Claim
22 words
A method according to claim 30 wherein the step of providing said assay material further comprises providing TMS bulked in said TEA-TPA.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-02-18 | 1983-08-20 | 1984-02-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-02-18 | 1987-08-20 | 1988-02-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-02-18 | 1991-08-20 | 1992-02-19 | $7700.00 | Window Closed / Lapsed |