US4833075A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Immobilized luciferase for the quantitative determination of ATP
Assignee: Centre National de la Recherche Scientifique
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Filing: 1988-04-01
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Granted: 1989-05-23
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Expires: 2008-04-01
Abstract
Abstract Luciferase is immobilized on a membrane prepared from an absorbent albuminoid such as procine skin gelatin. The membrane may be formed by spreading a solution of porcine skin gelatin on a support and after incubation and drying removing the resultant membrane from the support. Preferably, the membrane is crosslinked with a solution of glutaraldehyde, and luciferase is immobilized by saturating the membrane with luciferase in the presence of dithiothreitol. The immobilized luciferase is particularly suitable for the quantitative determination of adenosine triphosphoric acid (ATP).
Inventors
- 👤 Mookambeswaran Vijayalakshmi
- 👤 Rajgopal Sunanda
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 16:36
CPC Classifications
C; C12; C12N; C12N11/00; C12N11/02; C12Q; C12Q1/00; C12Q1/66
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
63 words
A process for producing a membrane having luciferase immobilized thereon, comprising the steps of, dissolving a porcine skin gelatin in a slightly acidic buffer and obtaining a homogenous membrane, cross-linking said homogenous membrane in a glutaraldehyde solution at a slightly acidic pH, and saturating the cross-linked membrane with luciferase in the presence of dithiothreitol to obtain a membrane which is saturated with luciferase.
Claim 2
Dependent Claim
63 words
A process for producing a membrane having luciferase immobilized thereon, comprising the steps of, dissolving a porcine skin gelatin in a slightly acidic buffer and obtaining a homogenous membrane, cross-linking said homogenous membrane in a glutaraldehyde solution at a slightly acidic pH, and saturating the cross-linked membrane with luciferase in the presence of dithiothreitol to obtain a membrane which is saturated with luciferase.
Claim 3
Dependent Claim
22 words
A process according to claim 1, wherein the slightly acid buffer is a solution of sodium phosphate having a pH of 6.8.
Claim 4
Dependent Claim
19 words
A process according to claim 1, wherein 6% by weight of porcine skin gelatin is dissolved in said buffer.
Claim 5
Dependent Claim
46 words
A process according to claim 1, wherein after dissolving said porcine skin gelatin, the resultant solution is incubated at between 25° and 50° C. for a short time, spread on a suitable support, dried to form a membrane, and the membrane is removed from the support.
Claim 6
Dependent Claim
17 words
A process according to claim 4, wherein incubation is carried out for one hour at 37° C.
Claim 7
Dependent Claim
15 words
A process according to claim 4, wherein the suitable support consists of a polycarbonate film.
Claim 8
Dependent Claim
15 words
A process according to claim 1, wherein the glutaraldehyde solution contains 0.5% by volume glutaraldehyde.
Claim 9
Dependent Claim
23 words
A process according to claim 7, wherein the pH of the said glutaraldehyde solution is the same as that of the said buffer.
Claim 10
Dependent Claim
15 words
A membrane on which luciferase is immobilized prepared according to the process of claim 1.
Claim 11
Dependent Claim
95 words
A process for quantitatively determining the presence of adenosine triphosphoric acid comprising the step of applying a material to be tested to a membrane on which luciferase is immobilized and measuring intensity of bioluminescence produced, said membrane containing immobilized luciferase being prepared by the steps of, dissolving a porcine skin gelatin in a slightly acidic buffer and obtaining a homogenous membrane, cross-linking said homogenous membrane in a glutaraldehyde solution at a slightly acidic pH, and saturating the cross-linked membrane with luciferase in the presence of dithiothreitol to obtain a membrane which is saturated with luciferase.
Claim 12
Dependent Claim
95 words
A process for quantitatively determining the presence of adenosine triphosphoric acid comprising the step of applying a material to be tested to a membrane on which luciferase is immobilized and measuring intensity of bioluminescence produced, said membrane containing immobilized luciferase being prepared by the steps of, dissolving a porcine skin gelatin in a slightly acidic buffer and obtaining a homogenous membrane, cross-linking said homogenous membrane in a glutaraldehyde solution at a slightly acidic pH, and saturating the cross-linked membrane with luciferase in the presence of dithiothreitol to obtain a membrane which is saturated with luciferase.
References Cited (Backward Citations)
US4601981A
Cited
US4411999A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-05-22 | 1992-11-21 | 1993-05-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-05-22 | 1996-11-21 | 1997-05-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-05-22 | 2000-11-21 | 2001-05-23 | $7700.00 | Window Closed / Lapsed |