US4434228A
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Immobilization of biological materials in condensed polyalkyleneimine polymers
Assignee: Genex Corp
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Filing: 1982-04-20
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Granted: 1984-02-28
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Expires: 2002-04-20
Abstract
Abstract Biological materials are immobilized within condensed polyalkyleneimine polymers. Condensation of the polymer is accomplished by bridging the amine groups of polyalkyleneimine polymer chains with a polycarboxylic acid, in the presence of a condensing agent. d
Inventors
- 👤 Wayne E. Swann
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 17:06
CPC Classifications
C; C12; C12P; C12P13/00; C12P13/04; C12P13/22; C12N; C12N11/00; C12N11/02; C12N11/08; C12N11/089; C12N11/098; C12P13/20
Claims Analysis
Total Claims: 39
Claim 1
Independent Claim
52 words
A method for immobilizing a biological material by preparing an insolubilized biological material composite, comprising condensing a polyalkyleneimine polymer with a condensing amount of a polycarboxylic acid in the presence of a condensing amount of a condensing agent, wherein said biological material is mixed with said polyalkyleneimine polymer during said condensation reaction.
Claim 2
Independent Claim
52 words
A method for immobilizing a biological material by preparing an insolubilized biological material composite, comprising condensing a polyalkyleneimine polymer with a condensing amount of a polycarboxylic acid in the presence of a condensing amount of a condensing agent, wherein said biological material is mixed with said polyalkyleneimine polymer during said condensation reaction.
Claim 3
Dependent Claim
21 words
The method of claim 1, wherein the polyalkyleneimine polymer is selected from the group consisting of polyethyleneimine, polypropyleneimine, polybutyleneimine and polypentyleneimine.
Claim 4
Dependent Claim
11 words
The method of claim 1, wherein the polyalkylenemine polymer is polyethyleneimine.
Claim 5
Dependent Claim
13 words
The method of claim 1, wherein the polycarboxylic acid is a dicarboxylic acid.
Claim 6
Dependent Claim
40 words
The method of claim 1, 2 or 3 wherein the polycarboxylic acid is selected from the group consisting of adipic, azelaic, 1,11-undecanedioic, 1,12-dodecanedioic, traumatic, pentadecanedioic, hexadecanedioic, sebacic, suberic, glutaric, malonic, pimellic, succinic, malic, maleic, glutamic, aspartic, oxalic, fumaric, and polyaspartic.
Claim 7
Dependent Claim
15 words
The method of claim 1, 2 or 3 wherein the polycarboxylic acid is succinic acid.
Claim 8
Dependent Claim
18 words
The method of claim 1 wherein the condensing agent is selected from the group consisting of N-ethyl-5-phenylisoxazolium-3-sulfonate, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline.
Claim 9
Dependent Claim
14 words
The method of claim 1 wherein the condensing agent is a carbodiimide condensing agent.
Claim 10
Dependent Claim
23 words
The method of claim 8, wherein the carbodiimide condensing agent is 1-ethyl-3,3-dimethylaminopropyl carbodiimide, dicyclohexyl carbodiimide, 1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide metho-p-toluene sulfonate, or a salt thereof.
Claim 11
Dependent Claim
17 words
The method of claim 1, wherein the carbodiimide condensing agent is 1-ethyl-3,3-dimethylaminopropyl carbodimide or a salt thereof.
Claim 12
Dependent Claim
16 words
The method of claim 5 wherein the condensing agent is 1-ethyl-3,3-dimethylaminopropyl carbodiimide or a salt thereof.
Claim 13
Dependent Claim
23 words
The method of claim 1, 2, 3 or 4, wherein the molar ratio of polyalkyleneimine to polycarboxylic is from about 1:20 to 1:0005.
Claim 14
Dependent Claim
27 words
The method of claim 12, wherein the condensing agent is employed in an amount ranging from about 0.2 to 3 times stoichiometric, relative to the polycarboxylic acid.
Claim 15
Dependent Claim
27 words
The method of claim 12, wherein the condensing agent is employed in an amount ranging from about 0.5 to 1.5 times stoichiometric relative to the polycarboxylic acid.
Claim 16
Dependent Claim
30 words
The method of claim 1, 2, 3 or 4 wherein the polyalkyleneimine and polycarboxylic acid are first mixed, then the biological material is added, then the condensing agent is added.
Claim 17
Dependent Claim
13 words
The method of claim 1, wherein the biological material immobilized is an enzyme.
Claim 18
Dependent Claim
13 words
The method of claim 1, wherein the biological material immobilized comprises whole cells.
Claim 19
Dependent Claim
16 words
The method of claim 17, wherein the whole cells are E. coli cells which contain aspartase.
Claim 20
Dependent Claim
11 words
The method of claim 9, wherein the biological material is aspartase.
Claim 21
Dependent Claim
12 words
The method of claim 1, wherein the biological material comprises phenylalanine ammonia-lyase.
Claim 22
Dependent Claim
12 words
The method of claim 1, wherein the biological material comprises tryptophan synthetase.
Claim 23
Dependent Claim
29 words
The method of claim 1, further comprising post treatment of the insolubilized biological material composite with an amine crosslinking agent to impart additional strength and stability to the composite.
Claim 24
Dependent Claim
27 words
The method of claim 22, wherein the amine crosslinking agent is selected from the group consisting of glutaric dialdehyde, diisocyanates, polyisocyanates, 2,4,6-trichloro-S-triazine, bisoxirane, bisimidate, divinylsulfone, and 1,5-difluoro-2,4-dinitrobenzene.
Claim 25
Dependent Claim
13 words
The method of claim 22, wherein the amine crosslinking agent is glutaric dialdehyde.
Claim 26
Dependent Claim
29 words
An insolubilized biological material composite comprising a biologically active material immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine chains are bridged by a polycarboxylic acid.
Claim 27
Dependent Claim
29 words
An insolubilized biological material composite comprising a biologically active material immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine chains are bridged by a polycarboxylic acid.
Claim 28
Dependent Claim
31 words
The composite of claim 25, wherein the biologically active material is selected from the group consisting of enzymes, antigens, antibodies, antibiotics, carbohydrates, coenzymes, plant cells, bacteria, yeast, fungi and tissue cultures.
Claim 29
Dependent Claim
21 words
The composite of claim 25, wherein the biologically active material is selected from the group consisting of enzymes and whole cells.
Claim 30
Dependent Claim
12 words
The composite of claim 25, wherein the biologically active material is aspartase.
Claim 31
Dependent Claim
18 words
The composite of claim 25, wherein the biological material are whole cells of E. coli which contain aspartase.
Claim 32
Dependent Claim
12 words
The composite of claim 25, wherein the biological material comprises phenylalanine ammonia-lyase.
Claim 33
Dependent Claim
12 words
The composite of claim 25, wherein the biological material comprises tryptophan synthetase.
Claim 34
Independent Claim
49 words
A method for producing aspartic acid, comprising contacting, under aspartic acid-producing conditions, a substrate, containing ammonium fumarate, with an insolubilized biological material composite of aspartase or aspartase-containing microbial cells immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine polymer chains are bridged by a polycarboxylic acid.
Claim 35
Independent Claim
49 words
A method for producing aspartic acid, comprising contacting, under aspartic acid-producing conditions, a substrate, containing ammonium fumarate, with an insolubilized biological material composite of aspartase or aspartase-containing microbial cells immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine polymer chains are bridged by a polycarboxylic acid.
Claim 36
Independent Claim
51 words
A method for producing phenylalanine, comprising contacting, under phenylalanine-producing conditions, a substrate, containing t-cinnamic acid and ammonia, with an insolubilized biological material composite of phenylalanine ammonia-lyase or phenylalanine ammonia-lyase-containing microbial cells immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine polymer chains are bridged by a polycarboxylic acid.
Claim 37
Independent Claim
51 words
A method for producing phenylalanine, comprising contacting, under phenylalanine-producing conditions, a substrate, containing t-cinnamic acid and ammonia, with an insolubilized biological material composite of phenylalanine ammonia-lyase or phenylalanine ammonia-lyase-containing microbial cells immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine polymer chains are bridged by a polycarboxylic acid.
Claim 38
Independent Claim
49 words
A method for producing tryptophan, comprising contacting, under tryptophan-producing conditions, a substrate, containing indole and serine, with an insolubilized biological material composite of tryptophan synthetase or tryptophan-synthetase-containing microbial cells immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine polymer chains are bridged by a polycarboxylic acid.
Claim 39
Independent Claim
49 words
A method for producing tryptophan, comprising contacting, under tryptophan-producing conditions, a substrate, containing indole and serine, with an insolubilized biological material composite of tryptophan synthetase or tryptophan-synthetase-containing microbial cells immobilized within a condensed polyalkyleneimine polymer, wherein amino groups on the polyalkyleneimine polymer chains are bridged by a polycarboxylic acid.
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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) | 1987-02-27 | 1987-08-29 | 1988-02-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-02-27 | 1991-08-29 | 1992-02-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-02-27 | 1995-08-29 | 1996-02-28 | $7700.00 | Window Closed / Lapsed |