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US4464468A
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Immobilization of active protein by cross-linking to inactive protein
Assignee: Bpifrance Financement SA
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Filing: 1982-04-16
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Granted: 1984-08-07
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Expires: 2002-04-16
Abstract
Abstract A solution of an active protein substance and an inactive protein substance is reacted with a cross-linking agent, optionally in the presence of an inert carrier, under cross-linking conditions to produce articles comprising both active and inactive protein substances. The active protein substance comprises up to about 20 percent, e.g. from 1 to 20 percent by weight, based on the final weight of the total protein substance, whereas the cross-linking agent comprises from 0.5 to 8 percent by weight, based on the weight of the total treated mixture. The obtained articles are in the form of a solution or a suspension in aqueous medium, in the form of a film, in the form of a membrane, in the form of a fabric, in the form of a porous material, or in the form of a mass, such as granules, pills or tablets.
Inventors
- 👤 Stratis Avrameas
- 👤 Georges Broun
- 👤 Eric Selegny
- 👤 Daniel Thomas
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 09:24
CPC Classifications
A; A61; A61K; A61K39/00; A61K39/395; A61K39/44; A61K35/00; A61K35/66; A61K35/74; A61K35/741; A61K35/742; A61K38/00; A61K38/16; A61K38/17; A61K38/36; A61K38/363; A61K38/38; A61K38/43; A61K38/44; A61K38/443; A61K38/46; A61K38/47; A61K38/48; A61K38/482; A61K38/4826; A61K38/51; C; C12; C12N; C12N11/00; C12N11/02; Y; Y10; Y10S; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/815; Y10S530/816
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
117 words
A Self-supporting open-cell foam or sponge consisting essentially of a homogenous distribution of active protein and inactive protein cross-linked together with glutaraldehyde having the following properties: (a) surface area--10 3 cm 2 /cm 3 or, when dry, 1 m 2 /g; (b) specific gravity--0.1 when dry (including open cells); (c) free volume--90 percent; (d) open cell sizes--from 1 to 200 microns; (e) capacity for water absorption--10 times its volume when starting with dry material; (f) absorption capacity--30 percent when considering only the insoluble phase; (g) elasticity (tension modulus)--0.4 to 0.7 kg/mm 2 ; (h) compression--0.2 to 0.9 kg/mm 2 , said active protein comprising up to 20 percent by weight of the total active and inactive proteins.
Claim 2
Dependent Claim
117 words
A Self-supporting open-cell foam or sponge consisting essentially of a homogenous distribution of active protein and inactive protein cross-linked together with glutaraldehyde having the following properties: (a) surface area--10 3 cm 2 /cm 3 or, when dry, 1 m 2 /g; (b) specific gravity--0.1 when dry (including open cells); (c) free volume--90 percent; (d) open cell sizes--from 1 to 200 microns; (e) capacity for water absorption--10 times its volume when starting with dry material; (f) absorption capacity--30 percent when considering only the insoluble phase; (g) elasticity (tension modulus)--0.4 to 0.7 kg/mm 2 ; (h) compression--0.2 to 0.9 kg/mm 2 , said active protein comprising up to 20 percent by weight of the total active and inactive proteins.
Claim 3
Dependent Claim
161 words
A process for producing an immobilized active protein in the form of a self-supporting flexible sheet or film or a self-supporting open-cell foam or sponge consisting essentially of mixing (a) active protein and (b) inactive protein simultaneously with (c) glutaraldehyde in a solvent-containing reaction medium to form a reaction mixture and reacting with active and inactive proteins with said glutaraldehyde under cross-linking conditions until the active protein is cross-linked with the inactive protein through bridges provided by the glutaraldehyde to form said sheet, film, foam or sponge, said active protein comprising up to 20 percent by weight of total active and inactive proteinic substance and said glutaraldehyde being in the range of from 0.5 to 8 percent by weight, based on the weight of total reaction mixture, said sheet or film being formed by spreading said reaction mixture on a surface before cross-linking and said foam or sponge being formed by freezing and slowly thawing said reaction mixture during said cross-linking.
Claim 4
Dependent Claim
161 words
A process for producing an immobilized active protein in the form of a self-supporting flexible sheet or film or a self-supporting open-cell foam or sponge consisting essentially of mixing (a) active protein and (b) inactive protein simultaneously with (c) glutaraldehyde in a solvent-containing reaction medium to form a reaction mixture and reacting with active and inactive proteins with said glutaraldehyde under cross-linking conditions until the active protein is cross-linked with the inactive protein through bridges provided by the glutaraldehyde to form said sheet, film, foam or sponge, said active protein comprising up to 20 percent by weight of total active and inactive proteinic substance and said glutaraldehyde being in the range of from 0.5 to 8 percent by weight, based on the weight of total reaction mixture, said sheet or film being formed by spreading said reaction mixture on a surface before cross-linking and said foam or sponge being formed by freezing and slowly thawing said reaction mixture during said cross-linking.
Claim 5
Dependent Claim
26 words
A process according to claim 2 wherein cross-linking is effected while reducing the amount of solvent in the reaction medium to form said sheet or film.
Claim 6
Dependent Claim
13 words
A process according to claim 2 wherein said foam or sponge is formed.
Claim 7
Dependent Claim
26 words
A process according to claim 2 wherein the inactive protein is an albumin and the amount of solvent in the reaction medium is reduced during cross-linking.
Claim 8
Dependent Claim
31 words
A process according to claim 2 wherein said sheet or film is formed, the inactive protein is gelatin and the amount of solvent in the reaction medium is reduced during cross-linking.
Claim 9
Dependent Claim
11 words
An immobilized active protein produced by the process of claim 2.
Claim 10
Dependent Claim
35 words
Immobilized active protein according to claim 7 wherein the active protein is a member selected from the group consisting of antigen, allergen, antibody, hormone, enzyme and proteinic part of a virus or of a cell.
Claim 11
Dependent Claim
27 words
Immobilized active protein according to claim 7 wherein the active protein is a member selected from the group consisting of oxidase, hydroxylase, isomerase, transferase, lyase and oxidoreductase.
Claim 12
Dependent Claim
26 words
Immobilized active protein according to claim 7 wherein the active protein is a member selected from the group consisting of pepsin, subtilisin, trypsin, chymotrypsin and papain.
Claim 13
Dependent Claim
55 words
Immobilized active protein according to claim 7 wherein the active protein is a member selected from the group consisting of glucose-oxidase, carbonic anhydrase, lipolytic hydrolase, amylolytic hydrolase, urease, asparaginase, uricase, peroxidase, catalase, phenylalanine-hydroxylase, galactose phosphate uridyl transferase, pronase, collagenase, keratinase, elastase, urate oxidase, tyrosine decarboxylase, hexokinase, phosphatase, L-amino-acid-oxidase, xanthine oxidase, decarboxylase, ribonuclease, α-amylase and β-galactosidase.
Claim 14
Dependent Claim
18 words
Immobilized active protein according to claim 7 wherein the active protein substantially retains both its activity and specificity.
Claim 15
Dependent Claim
28 words
Immobilized active protein according to claim 7 wherein the inactive protein is a member selected from the group consisting of albumin, plasma protein, ovalbumin, fibrinogen, gelatin and hemoglobin.
Claim 16
Dependent Claim
16 words
Immobilized active protein according to claim 7 wherein the inactive protein is that found in microbes.
Claim 17
Dependent Claim
13 words
Immobilized active protein according to claim 7 wherein the inactive protein is albumin.
Claim 18
Dependent Claim
33 words
Immobilized active protein according to claim 8 wherein the inactive protein is a member selected from the group consisting of albumin, plasma protein, ovalbumin, fibrinogen, gelatin, hemoglobin and inactive protein found in microbes.
Claim 19
Dependent Claim
33 words
Immobilized active protein according to claim 9 wherein the inactive protein is a member selected from the group consisting of albumin, plasma protein, ovalbumin, fibrinogen, gelatin, hemoglobin and inactive protein found in microbes.
Claim 20
Dependent Claim
33 words
Immobilized active protein according to claim 10 wherein the inactive protein is a member selected from the group consisting of albumin, plasma protein, ovalbumin fibrinogen, gelatin, hemoglobin and inactive protein found in microbes.
Claim 21
Dependent Claim
33 words
Immobilized active protein according to claim 11 wherein the inactive protein is a member selected from the group consisting of albumin, plasma protein, ovalbumin, fibrinogen, gelatin, hemoglobin and inactive protein found in microbes.
Claim 22
Dependent Claim
13 words
Immobilized active protein according to claim 7 wherein the inactive protein is gelatin.
Claim 23
Dependent Claim
14 words
Immobilized active protein according to claim 7 wherein the active protein is an antigen.
Claim 24
Dependent Claim
14 words
Immobilized active protein according to claim 7 wherein the active protein is an antibody.
Claim 25
Dependent Claim
14 words
Immobilized active protein according to claim 7 wherein the active protein is an allergen.
Claim 26
Dependent Claim
17 words
Immobilized active protein according to claim 7 wherein the active protein is a part of a virus.
Claim 27
Dependent Claim
16 words
Immobilized active protein according to claim 12 wherein the inactive protein is that of a microbe.
Claim 28
Dependent Claim
22 words
Immobilized active protein according to claim 7 wherein the active protein and the inactive protein are derived from a single impure source.
Claim 29
Dependent Claim
16 words
Immobilized active protein according to claim 26 wherein the single impure source is a microbe source.
Claim 30
Dependent Claim
22 words
A flexible sheet or film of immobilized active protein according to claim 1 which is free from a separate or distinct carrier.
Claim 31
Dependent Claim
25 words
A sheet or film of immobilized active protein according to claim 7 in combination with an inert non-proteinic carrier which imparts form and strength thereto.
Claim 32
Dependent Claim
45 words
A sheet or film of immobilized active protein according to claim 29 wherein the carrier is of a material selected from the group consisting of cellulose, regenerated cellulose, amylose, an alginate, a polysilane, polyvinyl alcohol, aminated paper, a polyacrylamide, silicone sheet, activated carbon and silk.
Claim 33
Dependent Claim
16 words
Immobilized active protein according to claim 7 in the form of an open-cell foam or sponge.
Claim 34
Dependent Claim
14 words
Immobilized active protein according to claim 7 wherein the active protein is hydrolytic enzyme.
Claim 35
Dependent Claim
14 words
Immobilized active protein according to claim 7 wherein the active protein is proteolytic hydrolase.
Claim 36
Dependent Claim
39 words
Immobilized active protein according to claim 8 wherein the active protein substantially retains both its activity and specificity and the inactive protein is a member selected from the group consisting of albumin, plasma protein, ovalbumin, fibrinogen, gelatin and hemoglobin.
References Cited (Backward Citations)
US3639558A
Cited
US4004979A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-08-07 | 1988-02-06 | 1988-08-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-08-07 | 1992-02-06 | 1992-08-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-08-07 | 1996-02-06 | 1996-08-07 | $7700.00 | Window Closed / Lapsed |