USRE32016E
Expired / Lapsed
US
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Analysis of lactic acid or lactate using lactate oxidase
Assignee: Clinical Diagnostic Systems Inc
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Filing: 1981-08-31
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Granted: 1985-10-29
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Expires: 2001-08-31
Abstract
Abstract Compositions and multilayer analytical elements comprising lactate oxidase which is substantially free of catalase and preferably produced by Streptococcus faecalis ATCC 12755 are provided for the quantitative analysis of lactic acid or lactate, especially in serum. The lactate oxidase catalyzes the reduction of lactic acid or lactate to pyruvate and hydrogen peroxide and the quantity of lactic acid or lactate is determined by detecting the amount of hydrogen peroxide produced. Preferably, the hydrogen peroxide is detected colorimetrically using a peroxidase-catalyzed detection system.
Inventors
- 👤 Theodore W. Esders
- 👤 Charles T. Goodhue
- 👤 Richard M. Schubert
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 08:19
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0006; C12Q; C12Q1/00; C12Q1/26
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
97 words
A composition for the assay of lactic acid or lactate, said composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity.
Claim 2
Dependent Claim
97 words
A composition for the assay of lactic acid or lactate, said composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity.
Claim 3
Dependent Claim
18 words
A composition as described in claim 1 wherein the lactate oxidase is obtained from Streptococcus faecalis ATCC 12755.
Claim 4
Dependent Claim
16 words
A composition as described in claim 1 wherein the substance having peroxidative activity is a peroxidase.
Claim 5
Dependent Claim
24 words
A composition as described in claim 1 wherein the material that undergoes a detectable change is an organic compound tha undergoes a visible change.
Claim 6
Dependent Claim
116 words
An element for the detection of lactic acid or lactate, the element comprising a spreading layer and a reagent layer in fluid contact under conditions of use and containing a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity.
Claim 7
Dependent Claim
116 words
An element for the detection of lactic acid or lactate, the element comprising a spreading layer and a reagent layer in fluid contact under conditions of use and containing a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity.
Claim 8
Dependent Claim
15 words
An element as described in claim 5 wherein said reagent layer contains said lactate oxidase.
Claim 9
Dependent Claim
40 words
An element as described in claim 5 wherein said reagent layer contains a composition comprising a substance having peroxidative activity and a material that undergoes a detectable change in the presence of hydrogen peroxide and said substance having peroxidative activity.
Claim 10
Dependent Claim
16 words
An element as described in claim 7 wherein said substance having peroxidative activity is a peroxidase.
Claim 11
Dependent Claim
24 words
An element as described in claim 7 wherein the material that undergoes a detectable change is an organic compound that undergoes a visible change.
Claim 12
Dependent Claim
18 words
An element as described in claim 5 wherein the lactate oxidase is obtained from Streptococcus faecalis ATCC 12755.
Claim 13
Dependent Claim
43 words
The element of claim 5 wherein said composition is contained in the reagent layer and further including a third layer, between the spreading layer and the reagent layer, for preventing the material which undergoes a detectable change from migrating into the spreading layer.
Claim 14
Dependent Claim
22 words
The element as described in claim 11 which further comprises a support, the reagent layer separating the support from the spreading layer.
Claim 15
Dependent Claim
16 words
An element as described in claim 11 wherein said substance having peroxidative activity is a peroxidase.
Claim 16
Dependent Claim
24 words
An element as described in claim 11 wherein the material that undergoes a detectable change is an organic compound that undergoes a visible change.
Claim 17
Dependent Claim
18 words
An element as described in claim 11 wherein the lactate oxidase is obtained from Streptococcus faecalis ATCC 12755.
Claim 18
Dependent Claim
68 words
An element for the assay of lactic acid or lactate comprising a spreading layer and a reagent layer in fluid contact under conditions of use and containing a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μmoles lactate per minute per mg of protein, (3) is soluble in water,
Claim 19
Dependent Claim
68 words
An element for the assay of lactic acid or lactate comprising a spreading layer and a reagent layer in fluid contact under conditions of use and containing a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μmoles lactate per minute per mg of protein, (3) is soluble in water,
Claim 20
Dependent Claim
87 words
(4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity, said components (B) and (C) being present in the reagent layer, the lactate oxidase being persent in a concentration of from about 50 to about 500 (B) being present in a concentration of from about 2000 to about 20,000 U/m 2 .
Claim 21
Dependent Claim
87 words
(4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity, said components (B) and (C) being present in the reagent layer, the lactate oxidase being persent in a concentration of from about 50 to about 500 (B) being present in a concentration of from about 2000 to about 20,000 U/m 2 .
Claim 22
Dependent Claim
39 words
The element as described in claim 16 wherein said element further comprises a third layer, located between the spreading layer and the reagent layer, for preventing the material that undergoes a detectable change from migrating into the spreading layer.
Claim 23
Dependent Claim
36 words
The element as described in claim 17 wherein the material that undergoes a detectable change is a triarylimidazole dye and (B) is present in a concentration of from about 3,000 to about 10,000 U/m 2 .
Claim 24
Dependent Claim
17 words
The element as described in claim 18 wherein the material that undergoes a detectable change is 2(3,5-dimethoxy-4-hydroxyphenyl)-4,5-bis(4-dimethylaminophenyl)imidazole.
Claim 25
Dependent Claim
36 words
The element as described in claim 17 wherein the material that undergoes a detectable change is a selfcoupling dye and (B) is present in a concentration of from about 3,000 to about 10,000 U/m 2 .
Claim 26
Dependent Claim
17 words
The element as described in claim 20 wherein the material that undergoes a detectable change is 4-isopropoxy-1-naphthol.
Claim 27
Dependent Claim
24 words
The element as described in claim 17 which further comprises a support, the reagent layer being nearer to the support than the spreading layer.
Claim 28
Dependent Claim
24 words
An element as described in claim 17 wherein the material that undergoes a detectable change is an organic compound that undergoes a visible change.
Claim 29
Dependent Claim
18 words
An element as described in claim 17 wherein the lactate oxidase is obtained from Streptococcus faecalis ATCC 12755.
Claim 30
Dependent Claim
106 words
An element for the detection of lactic acid or lactate comprising a matrix of absorbent material and imbibed therein a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water. (4) is substantially catalase-free and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity.
Claim 31
Dependent Claim
106 words
An element for the detection of lactic acid or lactate comprising a matrix of absorbent material and imbibed therein a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water. (4) is substantially catalase-free and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity.
Claim 32
Dependent Claim
18 words
An element as described in claim 25 wherein said lactate oxidase is obtained from Streptococcus faecalis ATCC 12755.
Claim 33
Independent Claim
195 words
A method for detecting lactic acid or lactate, acid method comprising: contacting in an aqueous medium a sample for analysis and a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity; and detecting said detectable change. .Iadd.28. An enzyme lactate oxidase having at least the following substrate specificity and enzyme action: substrate specificity: L-lactic acid enzyme action: catalyzes the reaction L-lactic acid+O 2 →pyruvic acid+H 2 O 2 wherein said enzyme lactate oxidase (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ mole lactate per minute per mg. of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for
Claim 34
Independent Claim
195 words
A method for detecting lactic acid or lactate, acid method comprising: contacting in an aqueous medium a sample for analysis and a composition comprising (A) a lactate oxidase which (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ moles lactate per minute per mg of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM, (B) a substance having peroxidative activity, and (C) a material that undergoes a detectable change in the presence of hydrogen peroxide and the substance having peroxidative activity; and detecting said detectable change. .Iadd.28. An enzyme lactate oxidase having at least the following substrate specificity and enzyme action: substrate specificity: L-lactic acid enzyme action: catalyzes the reaction L-lactic acid+O 2 →pyruvic acid+H 2 O 2 wherein said enzyme lactate oxidase (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.1μ mole lactate per minute per mg. of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for
Claim 35
Dependent Claim
116 words
lactate of at least about 0.2 mM. .Iaddend. .Iadd.29. A process for the manufacture of lactate oxidase having substrate specificity at least for L-lactic acid and catalyzing the reaction: L-lactic acid+O 2 →pyruvic acid+H 2 O 2 which comprises culturing Streptococcus faecalis ATCC 12755 in a nutrient culture medium, and isolating the lactate oxidase thus produced from the culture wherein said lactate oxidase (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.2μ mole lactate per minute per mg. of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM. .Iaddend.
Claim 36
Dependent Claim
116 words
lactate of at least about 0.2 mM. .Iaddend. .Iadd.29. A process for the manufacture of lactate oxidase having substrate specificity at least for L-lactic acid and catalyzing the reaction: L-lactic acid+O 2 →pyruvic acid+H 2 O 2 which comprises culturing Streptococcus faecalis ATCC 12755 in a nutrient culture medium, and isolating the lactate oxidase thus produced from the culture wherein said lactate oxidase (1) is capable of effecting the production of detectable hydrogen peroxide, (2) has a specific activity of at least about 0.2μ mole lactate per minute per mg. of protein, (3) is soluble in water, (4) is substantially catalase-free, and (5) has a K m for lactate of at least about 0.2 mM. .Iaddend.
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) | 1988-10-28 | 1989-04-29 | 1989-10-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-10-28 | 1993-04-29 | 1993-10-29 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-10-28 | 1997-04-29 | 1997-10-29 | $7700.00 | Window Closed / Lapsed |