US4235961A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method for photometric determination of the subunit B of creatine kinase and a reagent for carrying out the method
Assignee: Pfizer Health AB
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Filing: 1979-06-21
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Granted: 1980-11-25
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Expires: 1999-06-21
Abstract
Abstract A method for the photometric determination of the subunit B of creatine kinase (CK-B) in buffered water solution through reaction with creatine phosphate, ADP and the system luciferase/D-luciferin in the presence of a specific antibody inhibiting the subunit M of creatine kinase, comprises making the determination in the presence of a D-luciferin analog, such as L-luciferin, having a structure similar to D-luciferin and being a competitive inhibitor of the firefly luciferase reaction. Also disclosed is a reagent for carrying out the above method comprising creatine-phosphate, ADP, luciferase, D-luciferin, L-luciferin, CK-M inhibiting antibodies, diadenosinpentaphosphate, SH-reagent, magnesium salt and buffer as well as possibly stabilizers and/or complex forming reagent.
Inventors
- 👤 Arne T. Lundin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 18:20
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/66; C12Q1/48; C12Q1/50; Y; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
70 words
Method for photometric determination of the subunit B of creatine kinase (CK-B) in buffered water solution through reaction with creatine phosphate, ADP and the system luciferase/D-luciferin in the presence of a specific antibody inhibiting the subunit M of creatine kinase, characterized in that the determination is made in the presence of a D-luciferin analog having a structure similar to D-luciferin and being a competitive inhibitor of the firefly luciferase reaction.
Claim 2
Dependent Claim
70 words
Method for photometric determination of the subunit B of creatine kinase (CK-B) in buffered water solution through reaction with creatine phosphate, ADP and the system luciferase/D-luciferin in the presence of a specific antibody inhibiting the subunit M of creatine kinase, characterized in that the determination is made in the presence of a D-luciferin analog having a structure similar to D-luciferin and being a competitive inhibitor of the firefly luciferase reaction.
Claim 3
Dependent Claim
11 words
Method according to claim 1, characterized in that diadenosinpentaphosphate is added.
Claim 4
Dependent Claim
23 words
Method according to claims 1 or 2, characterized in that ADP in a quantity of 10 -5 to 5×10 -3 M is added.
Claim 5
Dependent Claim
21 words
Method according to claim 1, characterized in that the method is carried out at a pH value between 6.5 and 7.8.
Claim 6
Dependent Claim
19 words
Method according to claim 4, characterized in that a buffer concentration between 10 mM and 250 mM is used.
Claim 7
Dependent Claim
20 words
Method according to claim 1, characterized in that a thiol protecting reagent, preferably N-acetylcystein, glutathion, dithiotreitol or dithioerythrit, is added.
Claim 8
Dependent Claim
33 words
Reagent for carrying out a method for the photometric determination of the subunit B of creatine Kinase (CK-B), comprising creatine-phosphate, ADP, luciferase, D-luciferin, L-luciferin, CK-M inhibiting antibodies, diadenosinpentaphosphate, SH-reagent, magnesium salt and buffer.
Claim 9
Dependent Claim
33 words
Reagent for carrying out a method for the photometric determination of the subunit B of creatine Kinase (CK-B), comprising creatine-phosphate, ADP, luciferase, D-luciferin, L-luciferin, CK-M inhibiting antibodies, diadenosinpentaphosphate, SH-reagent, magnesium salt and buffer.
Claim 10
Dependent Claim
67 words
Reagent according to claim 7, characterized in that it comprises: ______________________________________
5 to 50 mM Creatine phosphate
1 × 10.sup.-5 to 5 × 10.sup.-3 M
ADP
0.5 to 20 μg/ml
Luciferase
25 to 500 μg/ml
D-luciferin
0.5 to 20 μg/ml
L-luciferin
CK-antibodies
5 × 10.sup.-8 to 5 × 10.sup.-6 M
diadenosinpentaphosphate
1 to 100 mM Thiolreagent
1 to 100 mM magnesiumacetate
10 to 250 mM Imidazol-acetate-buffer
______________________________________
Claim 11
Dependent Claim
69 words
Reagent according to claim 8, characterized in that it comprises: ______________________________________
8 to 15 mM creatine phosphate
1 × 10.sup.-5 to 5 × 10.sup.-4 M
ADP
1 to 3 μg/ml Luciferase
50 to 150 μg/ml
D-luciferin
1 to 10 μg/ml L-luciferin
CK-antibodies
1 × 10.sup.-7 to 1 × 10.sup.-6 M
Diadenosinpentaphosphate
2 to 50 mM Thiol reagent
5 to 20 mM magnesium acetate
60 to 120 mM Imidazol-acetate-buffer
______________________________________
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) | 1983-11-25 | 1984-05-26 | 1984-11-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-11-25 | 1988-05-26 | 1988-11-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-11-25 | 1992-05-26 | 1992-11-25 | $7700.00 | Window Closed / Lapsed |