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US4286057A Expired / Lapsed US ⏱️ 0.0 years remaining

Method and reagent for the determination of creatine kinase

Assignee: Roche Diagnostics GmbH
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Filing: 1980-02-28
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Granted: 1981-08-25
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Expires: 2000-02-28
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Abstract

Abstract Creatine kinase is determined by the reaction of creatine phosphate with adenosine diphosphate with the formation of adenosine triphosphate, reaction of the latter with luciferin and oxygen in the presence of luciferase and diadenosine pentaphosphate with the formation of oxyluciferin and adenosine monophosphate, and measurement of the light thereupon emitted, the reaction being performed with a saturated concentration of adenosine diphosphate and substrate, in the presence of 1 to 10 millimoles per liter of AMP at pH values of 5.8 to 7.5, with at least 50 units of luciferase per test. It is desirable to operate in the presence of diadenosine pentaphosphate or sodium fluoride and a sequestering agent.

Inventors

  • 👤 Karl Wulff
  • 👤 Fritz Stahler
  • 👤 Wolfgang Gruber

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-25 09:19
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/66; C12Q1/48; C12Q1/50; Y; Y10; Y10S; Y10S435/00; Y10S435/81

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 94 words
In a method for the determination of creatine kinase by the reaction of creatine phosphate with adenosinediphosphate with the formation of adenosine triphosphate, transformation of the latter with luciferin and oxygen in the presence of luciferase and diadenosine pentaphosphate with the formation of oxyluciferin and adenosine monophosphate, and measurement of the light emitted thereby, the improvement comprising performing the reaction at adenosine diphosphate substrate saturation concentration in the presence of 1 to 10 millimoles per liter of AMP, at pH values of 5.8 to 7.5, with at least 50 units per test of luciferase.
Claim 2
Dependent Claim 94 words
In a method for the determination of creatine kinase by the reaction of creatine phosphate with adenosinediphosphate with the formation of adenosine triphosphate, transformation of the latter with luciferin and oxygen in the presence of luciferase and diadenosine pentaphosphate with the formation of oxyluciferin and adenosine monophosphate, and measurement of the light emitted thereby, the improvement comprising performing the reaction at adenosine diphosphate substrate saturation concentration in the presence of 1 to 10 millimoles per liter of AMP, at pH values of 5.8 to 7.5, with at least 50 units per test of luciferase.
Claim 3
Dependent Claim 27 words
Improvement as claimed in claim 1 wherein 0.1 to 10 millimoles per liter of ADP and 1 to 100 micromoles per liter of diadenosine pentaphosphate are added.
Claim 4
Dependent Claim 45 words
Improvement as claimed in claim 2 wherein 0.5 to 2 millimoles per liter of ADP and 5 to 50 micromoles per liter of diadenosine pentaphosphate and 5 to 10 millimoles per liter of AMP are used and a pH of 6.2 to 7.2 is utilized.
Claim 5
Dependent Claim 18 words
Improvement as claimed in claim 1 wherein said determination is performed in the presence of a sequestering agent.
Claim 6
Dependent Claim 15 words
Improvement as claimed in claim 4 wherein the sequestering agent is ethylene diamine tetraacetic acid.
Claim 7
Dependent Claim 19 words
Improvement as claimed in claim 1 wherein said determination is performed in the presence of an organic sulfohydryl compound.
Claim 8
Dependent Claim 14 words
Improvement as claimed in claim 6 wherein said organic sulfohydryl compound is N-acetyl cysteine.
Claim 9
Dependent Claim 17 words
Improvement as claimed in claim 1 wherein the determination is performed in the presence of serum albumin.
Claim 10
Dependent Claim 96 words
Reagent for the determination of creatine kinase comprising 10 to 500 micromoles of luciferin, at least 50 units of luciferase, 5 to 250 millimoles of buffer substance, pH 5.8 to 7.5, 1 to 100 micromoles of diadenosine pentaphosphate, 1 to 10 millimoles of AMP, 5 to 50 millimoles of creatine phosphate, 0.1 to 10 millimoles of ADP, 1 to 100 millimoles of organic sulfohydryl compound, 0.1 to 5 millimoles of sequestering agent, 0.05 to 1 weight percent of bovine serum albumin, and 1 to 100 millimoles of magnesium ions for every one liter of test solution.
Claim 11
Dependent Claim 96 words
Reagent for the determination of creatine kinase comprising 10 to 500 micromoles of luciferin, at least 50 units of luciferase, 5 to 250 millimoles of buffer substance, pH 5.8 to 7.5, 1 to 100 micromoles of diadenosine pentaphosphate, 1 to 10 millimoles of AMP, 5 to 50 millimoles of creatine phosphate, 0.1 to 10 millimoles of ADP, 1 to 100 millimoles of organic sulfohydryl compound, 0.1 to 5 millimoles of sequestering agent, 0.05 to 1 weight percent of bovine serum albumin, and 1 to 100 millimoles of magnesium ions for every one liter of test solution.
Claim 12
Dependent Claim 95 words
Reagent as claimed in claim 9 comprising 15 to 50 micromoles of luciferin, 100 to 5×10 4 units of luciferase, 60 to 120 millimoles of buffer substance, 6.3 to 7.2 pH, 5 to 10 millimoles of AMP, 0.5 to 2 millimoles of ADP, 5 to 50 micromoles of diadenosine pentaphosphate, 20 to 40 millimoles of creatine phosphate, 2to 50 millimoles of organic sulfohydryl compound, 0.5 to 3 millimoles of sequestering agent, 0.05 to 0.4 weight percent of bovine serum albumin, and 5 to 20 millimoles of magnesium ions for every one liter of test solution.
Claim 13
Dependent Claim 16 words
Reagent as claimed in claim 9 also containing ethylene diamine tetraacetic acid as a sequestering agent.
Claim 14
Dependent Claim 13 words
Reagent as claimed in claim 9 containing N-acetyl cysteine as a sulfohydryl compound.
Claim 15
Dependent Claim 17 words
Reagent as claimed in claim 9 also containing imidazole acetate or hepes acetate as a buffer substance.

References Cited (Backward Citations)

US3575811A Cited
US4080265A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1984-08-24 1985-02-23 1985-08-25 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1988-08-24 1989-02-23 1989-08-25 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1992-08-24 1993-02-23 1993-08-25 $7700.00 Window Closed / Lapsed

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