US4371612A
Expired / Lapsed
US
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Immobilization of biological material with an acrylonitrile polymer
Assignee: Asahi Kasei Corp
•
Filing: 1980-12-01
•
Granted: 1983-02-01
•
Expires: 2000-12-01
Abstract
Abstract Biological material such as enzymes is immobilized by adsorption or covalent bonding to a micro-porous water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer. Covalent bonding may be carried out with a crosslinking or condensing agent. The amino groups may be introduced into the polymer by reduction of nitrile groups with lithium aluminum hydride in an inert non-solvent.
Inventors
- 👤 Kunio Matsumoto
- 👤 Rokuro Izumi
- 👤 Hideji Seijo
- 👤 Hiroyuki Mizuguchi
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 04:16
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0006; C08; C08F; C08F8/00; C08F8/30; C12N11/00; C12N11/02; C12N11/08; C12N11/082; C12N11/087; C12N9/14; C12N9/16; C12N9/78; C12N9/80; C12N9/84; C12P; C12P35/00; C12P35/02; C12P37/00; C12P37/06
Claims Analysis
Total Claims: 77
Claim 1
Independent Claim
87 words
An immobilized material comprising a biologically active enzyme bound by covalent bonding with a crosslinking agent or condensing agent onto a carrier which is a water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer and having a porous structure containing a number of micro-pores with an average size of from 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 2
Dependent Claim
87 words
An immobilized material comprising a biologically active enzyme bound by covalent bonding with a crosslinking agent or condensing agent onto a carrier which is a water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer and having a porous structure containing a number of micro-pores with an average size of from 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 3
Dependent Claim
30 words
An immobilized material according to claim 1, wherein the acrylonitrile polymer containing amino groups is shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 4
Dependent Claim
27 words
An immobilized material according to claim 2, wherein the acrylonitrile polymer contains amino groups in an amount which does not make the polymer substantially soluble in water.
Claim 5
Dependent Claim
32 words
An immobilized material according to claim 1, wherein the acrylonitrile polymer is a polymer of an acrylonitrile monomer or a copolymer of said monomer with a comonomer containing ethylenically unsaturated double bond.
Claim 6
Dependent Claim
17 words
An immobilized material according to claim 4, wherein the acrylonitrile monomer is acrylonitrile, methacrylonitrile, α-chloroacrylonitrile or cinnamnitrile.
Claim 7
Dependent Claim
48 words
An immobilized material according to claim 4, wherein the comonomer is a styrene monomer, an acrylic acid ester monomer, a conjugated diene, a halogenated olefin, a vinyl ether monomer, a vinyl ketone monomer, a vinyl ester monomer, an amide vinyl monomer, a basic monomer or a polyfunctional monomer.
Claim 8
Dependent Claim
34 words
An immobilized material according to claim 4, wherein the acrylonitrile polymer comprises polyacrylonitrile containing 90% or more of acrylonitrile shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 9
Dependent Claim
38 words
An immobilized material according to claim 4, wherein the acrylonitrile polymer is a copolymer consisting of acrylonitrile, divinyl benzene and vinyl ethyl benzene shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 10
Dependent Claim
34 words
An immobilized material according to claim 1, wherein said covalent bonding is effected directly through said crosslinking agent or condensing agent or through said crosslinking agent or condensing agent and through an intermediary spacer.
Claim 11
Dependent Claim
13 words
An immobilized material according to claim 9, wherein said crosslinking agent is glutaraldehyde.
Claim 12
Dependent Claim
31 words
An immobilized material according to claim 9, wherein said crosslinking agent is glutaraldehyde, crotonaldehyde, acrolein, hexamethylenediamine diisocyanate, hexamethylene-iso-thiocyanate, toluene diisocyanate, xylene diisocyanate, dimethyl adipiimidate, dimethyl suberiimidate, dimethyl-3,3'-dithio-bis-propionimidate or succinic acid anhydride.
Claim 13
Dependent Claim
13 words
An immobilized material according to claim 9, wherein said condensing agent is 1-ethyl-3-(3-dimethylamino-propyl)-carbodiimide.
Claim 14
Dependent Claim
14 words
An immobilized material according to claim 9, wherein said spacer is an alkylene diamine.
Claim 15
Dependent Claim
95 words
A process for preparing an immobilized material which comprises binding a carrier of a water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer with a biologically active enzyme by covalent bonding with a crosslinking agent or condensing agent in an inert medium; said polymer having a porous structure containing a number of micro-pores with an average size of 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 16
Dependent Claim
95 words
A process for preparing an immobilized material which comprises binding a carrier of a water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer with a biologically active enzyme by covalent bonding with a crosslinking agent or condensing agent in an inert medium; said polymer having a porous structure containing a number of micro-pores with an average size of 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 17
Dependent Claim
29 words
A process according to claim 14, wherein the acrylonitrile polymer containing amino groups is shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 18
Dependent Claim
29 words
A process according to claim 14 or claim 15, wherein the acrylonitrile polymer contains amino groups in an amount which does not make the polymer substantially soluble in water.
Claim 19
Dependent Claim
31 words
A process according to claim 14, wherein the acrylonitrile polymer is a polymer of an acrylonitrile monomer or a copolymer of said monomer with a comonomer containing ethylenically unsaturated double bond.
Claim 20
Dependent Claim
16 words
A process according to claim 17, wherein the acrylonitrile monomer is acrylonitrile, methacrylonitrile, α-chloroacrylonitrile or cinnamnitrile.
Claim 21
Dependent Claim
47 words
A process according to claim 17, wherein the comoner is a styrene monomer, an acrylic acid ester monomer, a conjugated diene, a halogenated olefin, a vinyl ether monomer, a vinyl ketone monomer, a vinyl ester monomer, an amide vinyl monomer, a basic monomer or a polyfunctional monomer.
Claim 22
Dependent Claim
33 words
A process according to claim 17, wherein the acrylonitrile polymer comprises polyacrylonitrile containing 90% or more of acrylonitrile shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 23
Dependent Claim
37 words
A process according to claim 17, wherein the acrylonitrile polymer is a copolymer consisting of acrylonitrile, divinyl benzene and vinyl ethyl benzene shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 24
Dependent Claim
32 words
A process according to claim 14, wherein covalent bonding is effected directly through said crosslinking agent or condensing agent or through said crosslinking agent or condensing agent and through an intermediary spacer.
Claim 25
Dependent Claim
30 words
A process according to claim 22, wherein the crosslinking agent is glutaraldehyde, crotonaldehyde, acrolein, hexamethylenediamine diisocyanate, hexamethylene-iso-thiocyanate, toluene diisocyanate, xylene diisocyanate, dimethyl adipiimidate, dimethyl suberiimidate, dimethyl-3,3'-dithio-bis-propionimidate or succinic acid anhydride.
Claim 26
Dependent Claim
12 words
A process according to claim 22, wherein the condensing agent is 1-ethyl-3-(4-dimethylaminopropyl)-carbodiimide.
Claim 27
Dependent Claim
13 words
A process according to claim 22, wherein the spacer is an alkylene diamine.
Claim 28
Independent Claim
135 words
A device for promoting an enzymatic reaction comprising a means containing an immobilized material having a biologically active enzyme bound by covalent bonding with a crosslinking agent or condensing agent to a carrier comprising a water-insoluble acrylonitrile polymer containing amino groups and having a porous structure having a number of micro-pores with an average pore size of 40-9000 A, said polymer containing 20 μM to 1000 μM of amino groups per gram of the polymer and said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert nonsolvent with lithium aluminum hydride, a storage means containing a liquid substrate on which said biologically active enzyme has an activity and means for transporting at least a portion of the substrate from said storage means to said immobilized material.
Claim 29
Independent Claim
135 words
A device for promoting an enzymatic reaction comprising a means containing an immobilized material having a biologically active enzyme bound by covalent bonding with a crosslinking agent or condensing agent to a carrier comprising a water-insoluble acrylonitrile polymer containing amino groups and having a porous structure having a number of micro-pores with an average pore size of 40-9000 A, said polymer containing 20 μM to 1000 μM of amino groups per gram of the polymer and said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert nonsolvent with lithium aluminum hydride, a storage means containing a liquid substrate on which said biologically active enzyme has an activity and means for transporting at least a portion of the substrate from said storage means to said immobilized material.
Claim 30
Dependent Claim
27 words
A device according to claim 26, wherein the acrylonitrile type polymer is shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 31
Dependent Claim
27 words
A device according to claim 27, wherein the acrylonitrile type polymer contains amino groups in an amount which does not make the polymer substantially soluble in water.
Claim 32
Dependent Claim
31 words
A device according to claim 26, wherein the acrylonitrile polymer is a polymer of an acrylonitrile monomer or a copolymer of said monomer with a comonomer containing ethylenically unsaturated double bond.
Claim 33
Dependent Claim
16 words
A device according to claim 29, wherein the acrylonitrile monomer is acrylonitrile, methacrylonitrile, α-chloroacrylonitrile or cinnamnitrile.
Claim 34
Dependent Claim
48 words
A device according to claim 29, wherein the comonomer is a styrene monomer, an acrylic acid ester monomer, a conjugated diene, a halogenated olefin, a vinyl ether monomer, a vinyl ketone type monomer, a vinyl ester monomer, an amide vinyl monomer, a basic monomer or a polyfunctional monomer.
Claim 35
Dependent Claim
33 words
A device according to claim 29, wherein the acrylonitrile polymer comprises polyacrylonitrile containing 90% or more of acrylonitrile shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 36
Dependent Claim
37 words
A device according to claim 29, wherein the acrylonitrile polymer is a copolymer consisting of acrylonitrile, divinyl benzene and vinyl ethyl benzene shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 37
Dependent Claim
23 words
A device according to claim 26, wherein the enzymatic reaction system is a system for detecting electrical change caused by said enzymatic reaction.
Claim 38
Dependent Claim
28 words
A device according to claim 34, wherein the electrical change caused by the enzymatic reaction system is a change caused by the oxidizing ability of the reaction product.
Claim 39
Dependent Claim
102 words
A device according to claim 26, wherein said means containing the immobilized material comprises a packed column having an inlet and an outlet through which the liquid substrate can pass and the storage means for the liquid substrate comprises a cooled storage tank equipped with a stirrer for effecting agitation of the liquid substrate contained therein and said means for transporting the liquid substrate comprises a conduit means and operatively associated pump means for withdrawing liquid substrate from the storage means and passing the liquid substrate through the packed column and thereafter returning the treated liquid substrate to the said storage means.
Claim 40
Dependent Claim
30 words
A device according to claim 36, wherein the storage tank is equipped with a pH meter for adjusting a change in pH within the liquid substrate contained in said tank.
Claim 41
Dependent Claim
34 words
A device according to claim 36, wherein said conduit means further includes a heat exchanger for maintaining the liquid substrate at a temperature which will promote reaction with the enzyme bound to the carrier.
Claim 42
Dependent Claim
33 words
A device according to claim 26, wherein said covalent bonding is effected directly through said crosslinking agent or condensing agent or through said crosslinking agent or condensing agent and through an intermediary spacer.
Claim 43
Dependent Claim
12 words
A device according to claim 39, wherein said crosslinking agent is glutaraldehyde.
Claim 44
Dependent Claim
30 words
A device according to claim 39, wherein the crosslinking agent is glutaraldehyde, crotonaldehyde, acrolein, hexamethylenediamine diisocyanate, hexamethylene-iso-thiocyanate, toluene diisocyanate, xylene diisocyanate, dimethyl adipiimidate, dimethyl suberiimidate, dimethyl-3,3'-dithio-bix-propionimidate or succinic acid anhydride.
Claim 45
Dependent Claim
12 words
A device according to claim 39, wherein said condensing agent is 1-ethyl-3-(3-dimethylamino-propyl)-carbodiimide.
Claim 46
Dependent Claim
13 words
A device according to claim 39, wherein said spacer is an alkylene diamine.
Claim 47
Independent Claim
116 words
A method for effecting an enzymatic reaction by use of an immobilized material, which comprises reacting an immobilized material comprising a biologically active enzyme bound to a carrier by covalent bonding, with a crosslinking agent or condensing agent, said carrier comprising an acrylonitrile polymer containing amino groups and having a porous structure having a number of micro-pores with an average pore size of 40-9000 A, said polymer containing 20 μM to 1000 μM of amino groups per gram of the polymer and acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride, with a substrate on which said biologically active enzyme has activity.
Claim 48
Independent Claim
116 words
A method for effecting an enzymatic reaction by use of an immobilized material, which comprises reacting an immobilized material comprising a biologically active enzyme bound to a carrier by covalent bonding, with a crosslinking agent or condensing agent, said carrier comprising an acrylonitrile polymer containing amino groups and having a porous structure having a number of micro-pores with an average pore size of 40-9000 A, said polymer containing 20 μM to 1000 μM of amino groups per gram of the polymer and acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride, with a substrate on which said biologically active enzyme has activity.
Claim 49
Dependent Claim
29 words
A method according to claim 44, wherein the acrylonitrile polymer containing amino groups is shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 50
Dependent Claim
30 words
A method according to claim 44 or claim 45, wherein the acrylonitrile type polymer contains amino groups in an amount which does not make the polymer substantially soluble in water.
Claim 51
Dependent Claim
32 words
A method according to claim 44, wherein the acrylonitrile type polymer is a polymer of an acrylonitrile monomer or a copolymer of said monomer with a comonomer containing ethylenically unsaturated double bond.
Claim 52
Dependent Claim
16 words
A method according to claim 47, wherein the acrylonitrile monomer is acrylonitrile, methacrylonitrile, α-chloroacrylonitrile or cinnamnitrile.
Claim 53
Dependent Claim
47 words
A method according to claim 47, wherein the comonomer is a styrene monomer, an acrylic acid ester monomer, a conjugated diene, a halogenated olefin, a vinyl ether monomer, a vinyl ketone monomer, a vinyl ester monomer, an amide vinyl monomer, a basic monomer or a polyfunctional monomer.
Claim 54
Dependent Claim
33 words
A method according to claim 47, wherein the acrylonitrile polymer comprises polyacrylonitrile containing 90% or more of acrylonitrile shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 55
Dependent Claim
37 words
A method according to claim 47, wherein the acrylonitrile polymer is a copolymer consisting of acrylonitrile, divinyl benzene and vinyl ethyl benzene shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 56
Dependent Claim
42 words
A method according to claim 44, wherein the enzyme is an oxidoreductase, hydrolase, transferase, lyase, isomerase or ligase and the substrate is a liquid medium comprising blood, urine, phospolipids, uric acid and glucose or a solution of a cephalosporin or penicillin compound.
Claim 57
Dependent Claim
33 words
A method according to claim 44, wherein said covalent bonding is effected directly through said crosslinking agent or condensing agent or through said crosslinking agent or condensing agent and through an intermediary spacer.
Claim 58
Dependent Claim
12 words
A method according to claim 53, wherein the crosslinking agent is glutaraldehyde.
Claim 59
Dependent Claim
30 words
A method according to claim 53, wherein the crosslinking agent is glutaraldehyde, crotonaldehyde, acrolein, hexamethylenediamine diisocyanate, hexamethylene-iso-thiocyanate, toluene diisocyanate, xylene diisocyanate, dimethyl adipiimidate, dimethyl suberiimidate, dimethyl-3,3'-dithio-bis-propionimidate or succinic acid anhydride.
Claim 60
Dependent Claim
12 words
A method according to claim 53, wherein the condensing agent is 1-ethyl-3-(3-dimethylamino-propyl)-carbodiimide.
Claim 61
Dependent Claim
13 words
A method according to claim 44, wherein the spacer is an alkylene diamine.
Claim 62
Dependent Claim
108 words
A process for preparation of 6-aminopenicillanic acid which comprises enzymatically deacylating a benzylpenicillin or alkaline salt thereof by using immobilized penicillin acylase bound to a carrier of water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer, with the penicillin acylase bound by covalent bonding with a crosslinking agent or condensing agent in an inert medium, said polymer having a porous structure containing a number of micro-pores with an average size of 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 63
Dependent Claim
108 words
A process for preparation of 6-aminopenicillanic acid which comprises enzymatically deacylating a benzylpenicillin or alkaline salt thereof by using immobilized penicillin acylase bound to a carrier of water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer, with the penicillin acylase bound by covalent bonding with a crosslinking agent or condensing agent in an inert medium, said polymer having a porous structure containing a number of micro-pores with an average size of 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 64
Dependent Claim
33 words
A process according to claim 58, wherein the acrylonitrile polymer comprises polyacrylonitrile containing 90% or more of acrylonitrile shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 65
Dependent Claim
23 words
A process according to claim 58, wherein said penicillin acylase is an enzyme preparation obtained from cultured filtrate of penicillin acylase producing microorganisms.
Claim 66
Dependent Claim
19 words
A process according to claim 60, wherein the penicillin acylase producing microorganism is a strain belonging to genus Bacillus.
Claim 67
Dependent Claim
15 words
A process according to claim 61, wherein said strain is Bacillus megaterium B-400 (FERM-P 748).
Claim 68
Dependent Claim
12 words
A process according to claim 58, wherein said crosslinking agent is glutaraldehyde.
Claim 69
Dependent Claim
18 words
A process according to claim 58, wherein said micro-pores have an average size of 100 to 2000 A.
Claim 70
Dependent Claim
105 words
A process for preparation of 7-amino-desacetoxy cephalosporanic acid which comprises enzymatically deacylating a 7-phenylacetamidodesacetoxy cephalosporanic acid by using immobilized acylase bound to a carrier of water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer, with the acylase bound by covalent bonding with a crosslinking agent or condensing agent in an inert medium, said polymer having a porous structure containing a number of micro-pores with an average size of 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 71
Dependent Claim
105 words
A process for preparation of 7-amino-desacetoxy cephalosporanic acid which comprises enzymatically deacylating a 7-phenylacetamidodesacetoxy cephalosporanic acid by using immobilized acylase bound to a carrier of water-insoluble acrylonitrile polymer containing from 20 μM to 1000 μM of amino groups per gram of the polymer, with the acylase bound by covalent bonding with a crosslinking agent or condensing agent in an inert medium, said polymer having a porous structure containing a number of micro-pores with an average size of 40 to 9000 A, said acrylonitrile polymer being produced by effecting reduction of an acrylonitrile polymer having said porous structure in an inert non-solvent with lithium aluminum hydride.
Claim 72
Dependent Claim
33 words
A process according to claim 65, wherein the acrylonitrile polymer comprises polyacrylonitrile containing 90% or more of acrylonitrile shaped in the form of a fiber, a particle, a membrane or a hollow filament.
Claim 73
Dependent Claim
21 words
A process according to claim 65, wherein said acylase is an enzyme preparation obtained from cultured filtrate of acylase producing microorganisms.
Claim 74
Dependent Claim
18 words
A process according to claim 67, wherein the acylase producing microorganism is a strain belonging to genus Bacillus.
Claim 75
Dependent Claim
15 words
A process according to claim 68, wherein said strain is Bacillus megaterium B-400 (FERM-P 748).
Claim 76
Dependent Claim
12 words
A process according to claim 65, wherein said crosslinking agent is glutaraldehyde.
Claim 77
Dependent Claim
18 words
A process according to claim 65, wherein said micropores have an average size of 100 to 2000 A.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-01-31 | 1986-08-02 | 1987-02-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-01-31 | 1990-08-02 | 1991-02-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-01-31 | 1994-08-02 | 1995-02-01 | $7700.00 | Window Closed / Lapsed |