US4539294A
Expired / Lapsed
US
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Immobilization of proteins on polymeric supports
Assignee: Akzona Inc
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Filing: 1982-09-30
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Granted: 1985-09-03
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Expires: 2002-09-30
Abstract
Abstract A protein is immobilized by a simple and effective method on a porous polymeric support by a first soaking in a dilute long-chain cationic solution and a second soaking in a dilute aqueous protein solution. The long chain cationic is preferably a nitrogen compound such as a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms. A preferred diamine is N-coco-1,3-diamino-propane. The protein may be an enzyme or hormone. A preferred enzyme is catalase or a two enzyme system of catalase and glucose oxidase.
Inventors
- 👤 Lincoln D. Metcalfe
- 👤 Dieter Frank
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 11:01
CPC Classifications
C; C12; C12N; C12N11/00; C12N11/02; C12N11/08; C12N11/082; C12N11/14; C12N11/18; Y; Y10; Y10S; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/815
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
82 words
A method of immobilizing a protein on a porous polymeric support, comprising a first soaking step in which said support is placed in a dilute solution of a long-chain cationic comprising a nitrogen compound having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent, said support being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a dilute aqueous solution of said protein.
Claim 2
Independent Claim
82 words
A method of immobilizing a protein on a porous polymeric support, comprising a first soaking step in which said support is placed in a dilute solution of a long-chain cationic comprising a nitrogen compound having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent, said support being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a dilute aqueous solution of said protein.
Claim 3
Dependent Claim
45 words
The method as set forth in claim 1, wherein said support is placed in said dilute solution of a cationic in a solvent for between 5 and 60 minutes and wherein said support is soaked for more than four hours in said second soaking step.
Claim 4
Dependent Claim
14 words
The method as set forth in claim 1, wherein said protein is a hormone.
Claim 5
Independent Claim
19 words
The method as set forth in claims 1, 2, or 3, wherein said long-chain cationic is the diamine N-coco-1,3-diaminopropane.
Claim 6
Dependent Claim
15 words
The method as set forth in claim 3, wherein said hormone is human chorionic gonadotropin.
Claim 7
Independent Claim
100 words
A method of immobilizing a hormone on a support, comprising a first soaking step in which said support is placed in a dilute solution of a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent for said diamine for between 5 and 60 minutes, said diamine solution being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a aqueous solution of said hormone, said aqueous solution having a concentration of said hormone of from 0.002% (w/v) to 0.2% (w/v).
Claim 8
Independent Claim
100 words
A method of immobilizing a hormone on a support, comprising a first soaking step in which said support is placed in a dilute solution of a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent for said diamine for between 5 and 60 minutes, said diamine solution being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a aqueous solution of said hormone, said aqueous solution having a concentration of said hormone of from 0.002% (w/v) to 0.2% (w/v).
Claim 9
Independent Claim
82 words
A method of immobilizing an enzyme on a porous polymeric support, comprising a first soaking step in which said support is placed in a dilute solution of a long-chain cationic comprising a nitrogen compound having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent, said support being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a dilute aqueous solution of said enzyme.
Claim 10
Independent Claim
82 words
A method of immobilizing an enzyme on a porous polymeric support, comprising a first soaking step in which said support is placed in a dilute solution of a long-chain cationic comprising a nitrogen compound having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent, said support being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a dilute aqueous solution of said enzyme.
Claim 11
Dependent Claim
46 words
The method as set forth in claim 7, wherein said support is placed in said dilute solution of said long-chain cationic in a solvent for between 10 and 60 minutes and wherein said support is soaked for more than four hours in said second soaking step.
Claim 12
Dependent Claim
13 words
The method as set forth in claim 8, wherein said solvent is acetone.
Claim 13
Independent Claim
17 words
The method as set forth in claims 7, 8, or 9, wherein said long-chain cationic is N-coco-1,3-diaminopropane.
Claim 14
Dependent Claim
21 words
The method as set forth in claim 7, wherein said enzyme is a two enzyme system of catalase and glucose oxidase.
Claim 15
Dependent Claim
21 words
The method as set forth in claim 8, wherein said enzyme is a two enzyme system of catalase and glucose oxidase.
Claim 16
Dependent Claim
21 words
The method as set forth in claim 9, wherein said enzyme is a two enzyme system of catalase and glucose oxidase.
Claim 17
Dependent Claim
21 words
The method as set forth in claim 10, wherein said enzyme is a two enzyme system of catalase and glucose oxidase.
Claim 18
Independent Claim
125 words
A method of immobilizing catalase on a porous polymeric support, comprising a pretreatment step in which said support is placed in a dilute solution of a metal salt in a salt solvent for between 5 and 10 minutes, rinsing said support with said salt solvent or another salt solvent, a first soaking step in which said support is placed in a dilute solution of a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent for said diamine for between 10 and 60 minutes, said diamine solution being gently agitated during said first soaking step, and a second soaking step in which said support is soaked with agitation in a 0.8% (w/v) aqueous catalase solution.
Claim 19
Independent Claim
125 words
A method of immobilizing catalase on a porous polymeric support, comprising a pretreatment step in which said support is placed in a dilute solution of a metal salt in a salt solvent for between 5 and 10 minutes, rinsing said support with said salt solvent or another salt solvent, a first soaking step in which said support is placed in a dilute solution of a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent for said diamine for between 10 and 60 minutes, said diamine solution being gently agitated during said first soaking step, and a second soaking step in which said support is soaked with agitation in a 0.8% (w/v) aqueous catalase solution.
Claim 20
Dependent Claim
14 words
The method as set forth in claim 15, wherein said salt solvent is acetone.
Claim 21
Dependent Claim
14 words
The method as set forth in claim 15, wherein said amine solvent is acetone.
Claim 22
Dependent Claim
14 words
The method as set forth in claim 16, wherein said amine solvent is acetone.
Claim 23
Dependent Claim
18 words
The method as set forth in claim 18, wherein said metal salt is SnCl 2 .2H 2 O.
Claim 24
Dependent Claim
18 words
The method as set forth in claim 18, wherein said metal salt is FeCl 2 .2H 2 O.
Claim 25
Independent Claim
15 words
The method as set forth in claims 19 or 20, wherein said diamine is N-coco-1,3-diaminopropane.
Claim 26
Dependent Claim
13 words
The method as set forth in claim 21, wherein said enzyme is catalase.
Claim 27
Independent Claim
111 words
A method of immobilizing catalase on a porous polymeric film support, comprising a pretreatment in which said support is placed in a 1% (w/v) metal salt solution of SnCl 2 .2H 2 O in acetone; removing said support from said metal salt solution after five minutes and rinsing said support with acetone; a first soaking step in which said support is immersed for 60 minutes in a 1% (w/v) diamine solution of N-coco-1,3-diaminopropane in chloroform; removing said support from said diamine solution and wetting said support with isopropanol; and a second soaking step in which said support is immersed for more than four hours in a 0.8% (v/v) aqueous catalase solution.
Claim 28
Independent Claim
111 words
A method of immobilizing catalase on a porous polymeric film support, comprising a pretreatment in which said support is placed in a 1% (w/v) metal salt solution of SnCl 2 .2H 2 O in acetone; removing said support from said metal salt solution after five minutes and rinsing said support with acetone; a first soaking step in which said support is immersed for 60 minutes in a 1% (w/v) diamine solution of N-coco-1,3-diaminopropane in chloroform; removing said support from said diamine solution and wetting said support with isopropanol; and a second soaking step in which said support is immersed for more than four hours in a 0.8% (v/v) aqueous catalase solution.
Claim 29
Independent Claim
82 words
A method for immobilizing glucose oxidase and catalase on a porous polymer powder support, comprising: a first soaking step in which said support is immersed for 10 minutes in a 1% (w/v) diamine solution of N-coco-1,3-diaminopropane in chloroform; removing said support from said diamine solution and wetting said support; and a second soaking step in which said support is immersed for more than four hours in a dilute buffer solution of 2.5% (v/v) glucose oxidase and 0.5% (v/v) catalase in deionized water.
Claim 30
Independent Claim
82 words
A method for immobilizing glucose oxidase and catalase on a porous polymer powder support, comprising: a first soaking step in which said support is immersed for 10 minutes in a 1% (w/v) diamine solution of N-coco-1,3-diaminopropane in chloroform; removing said support from said diamine solution and wetting said support; and a second soaking step in which said support is immersed for more than four hours in a dilute buffer solution of 2.5% (v/v) glucose oxidase and 0.5% (v/v) catalase in deionized water.
Claim 31
Independent Claim
29 words
The method as set forth in claims 1, 2, 3 or 7 wherein said long-chain cationic is selected from the group consisting of amines, diamines, and quaternary ammonium compounds.
Claim 32
Dependent Claim
15 words
The method as set forth in claim 25, wherein said long-chain cationic is a diamine.
Claim 33
Independent Claim
18 words
The method as set forth in claims 1, 6, 15, 23 or 24, wherein said support comprises polypropylene.
Claim 34
Dependent Claim
13 words
The method as set forth in claim 10, wherein said support comprises polypropylene.
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) | 1988-09-02 | 1989-03-04 | 1989-09-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-09-02 | 1993-03-04 | 1993-09-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-09-02 | 1997-03-04 | 1997-09-03 | $7700.00 | Window Closed / Lapsed |