US4865984A
Expired / Lapsed
US
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Dynamic continuous flow enzyme reactor
Assignee: Icahn School of Medicine at Mount Sinai
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Filing: 1988-02-08
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Granted: 1989-09-12
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Expires: 2008-02-08
Abstract
Abstract A dynamic continuous flow enzyme reactor and method for carrying out and analyzing phospholipid-dependent enzyme reactions are provided. The enzyme reactor is a tubular member preferably a capillary tube, coated on its inner surface with a planar phospholipid bilayer membrane, optionally and preferably containing an enzyme or enzyme cofactor. The reactor is connectable at one end to a means for delivering fluid flow reagents to the reactor and is connectable at a second end to a means for analyzing an effluent exiting from the reactor. Preferably the enzyme cofactor in the phospholipid membrane is purified tissue factor and the fluid reagents are inactive blood clotting factors which become activated upon interaction with the phospholipid membrane through a tissue factor-mediated reaction.
Inventors
- 👤 Yale Nemerson
- 👤 Vincent Turitto
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 14:56
CPC Classifications
C; C12; C12M; C12M21/00; C12M21/18; C12M23/00; C12M23/02; C12M23/06; C12Q; C12Q1/00; C12Q1/56; G; G01; G01N; G01N2333/00; G01N2333/90; G01N2333/914; G01N2333/948; G01N2333/95; G01N2333/964; G01N2333/96425; G01N2333/96427; G01N2333/9643; G01N2333/96433; G01N2333/96441; G01N2333/96444; G01N2333/96447; G01N2333/96463; G01N2333/974; G01N2405/00; G01N2405/04
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
63 words
A dynamic continuous flow enzyme reactor for carrying out and analyzing phospholipid-dependent enzymatic reactions which comprises a tubular housing coated on its inner surface with a planar phospholipid bilayer membrane, the housing being connectable at one opening to a means for delivering fluid flow reagents thereto and connectable at a second opening to a means for collecting and analyzing an effluent fluid therefrom.
Claim 2
Dependent Claim
63 words
A dynamic continuous flow enzyme reactor for carrying out and analyzing phospholipid-dependent enzymatic reactions which comprises a tubular housing coated on its inner surface with a planar phospholipid bilayer membrane, the housing being connectable at one opening to a means for delivering fluid flow reagents thereto and connectable at a second opening to a means for collecting and analyzing an effluent fluid therefrom.
Claim 3
Dependent Claim
13 words
The enzyme reactor of claim 1 wherein the housing is a capillary tube.
Claim 4
Dependent Claim
32 words
The enzyme reactor of claim 1 wherein the housing is a capillary tube having an inner diameter of about 0.1 to 1.1 mm and a length of about 1.0 to 15 cm.
Claim 5
Dependent Claim
26 words
The enzyme reactor of claim 1 wherein the phospholipid membrane comprises at least one additional component selected from the group consisting of enzymes and enzyme cofactors.
Claim 6
Dependent Claim
12 words
The enzyme reactor of claim 4 wherein the cofactor is tissue factor.
Claim 7
Dependent Claim
28 words
The enzyme reactor of claim 5 wherein the phospholipid membrane contains tissue factor in a ratio of approximately 1-10 moles of tissue factor per 100,000 moles of phospholipid.
Claim 8
Dependent Claim
15 words
The enzyme reactor of claim 1 wherein the phospholipid membrane comprises neutral and acidic phospholipids.
Claim 9
Dependent Claim
14 words
The enzyme reactor of claim 1 wherein the phospholipid membrane comprises phosphatidylcholine and phosphatidylserine.
Claim 10
Dependent Claim
19 words
The enzyme reactor of claim 1 wherein the phospholipid membrane comprises a mixture of 60-100% phosphatidylcholine and 0-40% phosphatidylserine.
Claim 11
Dependent Claim
19 words
The enzyme reactor of claim 1 wherein the phospholipid membrane comprises a mixture of 70% phosphatidylcholine and 30% phosphatidylserine.
Claim 12
Independent Claim
145 words
A method for continuously carrying out and measuring dynamic enzyme reactions comprising: (a) delivering reagents for performing an enzymatic reaction at a defined flow rate to an inlet in a dynamic continuous enzyme flow reactor comprising a tubular housing which is coated on its inner surface with a planar phospholipid bilayer membrane, wherein the reagents and membrane components are separately inactive for performing the enzymatic reaction, but together yield an enzymatically active system; (b) pumping the reagents through the enzyme reactor at a constant and defined flow rate so that an enzyme reaction mediated by contact of the reagents with the phospholipid membrane takes place in the enzyme reactor; (c) collecting a product of the enzyme reaction in an effluent from an exit in the enzyme reactor; and (d) measuring the amount of product formed during the enzyme reaction by means of a suitable assay.
Claim 13
Independent Claim
145 words
A method for continuously carrying out and measuring dynamic enzyme reactions comprising: (a) delivering reagents for performing an enzymatic reaction at a defined flow rate to an inlet in a dynamic continuous enzyme flow reactor comprising a tubular housing which is coated on its inner surface with a planar phospholipid bilayer membrane, wherein the reagents and membrane components are separately inactive for performing the enzymatic reaction, but together yield an enzymatically active system; (b) pumping the reagents through the enzyme reactor at a constant and defined flow rate so that an enzyme reaction mediated by contact of the reagents with the phospholipid membrane takes place in the enzyme reactor; (c) collecting a product of the enzyme reaction in an effluent from an exit in the enzyme reactor; and (d) measuring the amount of product formed during the enzyme reaction by means of a suitable assay.
Claim 14
Dependent Claim
26 words
Method according to claim 11 wherein the phospholipid membrane further comprises at least one additional component selected from the group consisting of enzymes and enzyme cofactors.
Claim 15
Dependent Claim
11 words
Method according to claim 12 wherein the cofactor is tissue factor.
Claim 16
Dependent Claim
15 words
Method according to claim 13 wherein the reagents comprise blood clotting factors and calcium ions.
Claim 17
Dependent Claim
52 words
Method according to claim 14 wherein the blood clotting factors comprise a blood clotting factor selected from the group consisting of factor VII and factor VII a , together with at least one factor selected from the group consisting of factor IX, factor X, factor V, factor VIII, prothrombin and whole plasma.
Claim 18
Dependent Claim
16 words
Method according to claim 14 wherein the blood clotting factors are factor VII and factor IX.
Claim 19
Dependent Claim
17 words
Method according to claim 14 wherein the blood clotting factors are factor VII a and factor IX.
Claim 20
Dependent Claim
16 words
Method according to claim 14 wherein the blood clotting factors are factor VII and factor X.
Claim 21
Dependent Claim
17 words
Method according to claim 14 wherein the blood clotting factors are factor VII a and factor X.
Claim 22
Dependent Claim
19 words
Method according to claim 14 wherein the blood clotting factors are factor VII, factor X, factor V and prothrombin.
Claim 23
Dependent Claim
21 words
Method according to claim 14 wherein the blood clotting factors are factor VII a , factor X, factor V and prothrombin.
Claim 24
Dependent Claim
27 words
Method according to claim 11 wherein the assay for measuring the amount of product is selected from the group consisting of radioassays, chromogenic assays and fluorogenic assays.
Claim 25
Dependent Claim
30 words
Method according to claim 11 further comprising adding a chromogenic or fluorogenic substrate specific for the product of the reaction to the effluent before measuring the amount of product formed.
Claim 26
Dependent Claim
32 words
Method according to claim 11 further comprising splitting the product effluent from the enzyme reactor such that the amount of more than one product formed in the enzyme reactor can be measured.
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) | 1992-09-11 | 1993-03-13 | 1993-09-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-09-11 | 1997-03-13 | 1997-09-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-09-11 | 2001-03-13 | 2001-09-12 | $7700.00 | Window Closed / Lapsed |