Patent EP0088818A1 successfully refreshed from Google Patents.
EP0088818A1
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EP
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Support matrix for amino-containing biologically active substances
Assignee: Hexion Canada Inc
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Filing: 1982-03-17
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Granted: —
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Expires: 2002-03-17
Abstract
Abstract An improved immobilized enzyme composite is prepared by subjecting an inorganic carrier material which is essentially chemically inert however hydrolisable by a strong base or a strong acid to give raise to multiple OH groups and which is sufficiently porous or finely divided to give access to the base and the acid and further dissolved reactants to all parts thereof, especially a siliceous carrier to an initial treatment with alkali, then acid, then reacting the carrier with an organosilane. The organosilane may then be coupled through a covalent coupling agent with an enzyme. The alkali-acid treatment provides an exceptionally high number of hydroxyl groups per unit volume, so that the treated carrier is especially useful in the preparation of high performance immobilized enzyme compositions, particularly by multi-layering immobilization, providing a very high amount of enzyme activity per unit volume. Multi-layering is accomplished by successive steps of bonding a difunctional covalent coupling agent to a previously immobilized enzyme layer, then bonding an additional layer of enzyme to the coupling agent. This may be repeated to apply as many layers as desired.
Inventors
- 👤 Ho Guan Huei
- 👤 Liao Chiang-Chang
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 17:01
CPC Classifications
B; B01; B01J; B01J20/00; B01J20/30; B01J20/32; B01J20/3202; B01J20/3204; B01J20/02; B01J20/10; B01J20/103; B01J20/3085; B01J20/3214; B01J20/3217; B01J20/3219; B01J20/3231; B01J20/3242; B01J20/3244; B01J20/3246; B01J20/3257; B01J20/3259; B01J20/3261; B01J20/3268; B01J20/3272; B01J20/3274; B01J20/328; B01J20/3282; B01J20/3289; C; C12; C12N; C12N11/00; C12N11/14; B01J2220/00; B01J2220/50; B01J2220/58
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
136 words
A process for reacting an inorganic particulate carrier material which is hydrolisable by a strong base or a strong acid to form OH groups, especially a finely divided siliceous inorganic carrier material to activate its surface area with hydroxyl groups, comprising reacting the material, especially the siliceous carrier material at a pH of at least about 10 with a strong basic solution in which said carrier material retains its particulate form, for a sufficient time to generate surface groups on the particles of the carrier material of the type SiO - M + , where M is a metallic ion, an ammonium ion, or represents a mixture thereof, and then replacing the M with H by reacting the carrier material with a strongly acid solution, so that the carrier suface has reactive hydroxyl groups disposed thereon.
Claim 2
Dependent Claim
136 words
A process for reacting an inorganic particulate carrier material which is hydrolisable by a strong base or a strong acid to form OH groups, especially a finely divided siliceous inorganic carrier material to activate its surface area with hydroxyl groups, comprising reacting the material, especially the siliceous carrier material at a pH of at least about 10 with a strong basic solution in which said carrier material retains its particulate form, for a sufficient time to generate surface groups on the particles of the carrier material of the type SiO - M + , where M is a metallic ion, an ammonium ion, or represents a mixture thereof, and then replacing the M with H by reacting the carrier material with a strongly acid solution, so that the carrier suface has reactive hydroxyl groups disposed thereon.
Claim 3
Dependent Claim
16 words
2 . The process of claim 1 wherein the siliceous inorganic carrier material is silica gel.
Claim 4
Dependent Claim
72 words
The process of claim 1 or 2 wherein the strongly basic solution is a solution of an alkali metal hydroxide having a concentration of at least 0,1% by weight, or the equivalent, preferably an aqueous solution of an alkali metal hydroxide and a concentration up to about 5% by weight, the reaction temperature is at least 20°C and the reaction in the strongly basic solution is continued for at least 30 minutes.
Claim 5
Dependent Claim
42 words
The process of claim 3, wherein the reaction time is in the range from about 30 minutes to about 2 hours, after which the carrier material is treated with a strong acid to replace the M with H,preferably with a mineral acid.
Claim 6
Dependent Claim
135 words
A process for developing a high .density of reactive hydroxyl groups on the surfaces of particles of finely divided silica gel, comprising reacting silica gel with an aqueous solution of a strongly basic material selected from the group consisting of alkali metal hydroxides, alkali metal carbonates, and mixtures thereof, at a pH of at least about 10, at a temperature in the range from about 20°C to about 45°C, for a period of time from about 30 minutes to about 2 hours, during which reaction the silica gel retains its particulate form, seperating the alkali-reacted, particulate silica g el from the basic solution, and then reacting the said alkali-reacted, silicia gel particles with an aqueous mineral acid solution to remove metal ion from the gel and to activate the gel sufaces with reactive hydroxyl groups.
Claim 7
Dependent Claim
135 words
A process for developing a high .density of reactive hydroxyl groups on the surfaces of particles of finely divided silica gel, comprising reacting silica gel with an aqueous solution of a strongly basic material selected from the group consisting of alkali metal hydroxides, alkali metal carbonates, and mixtures thereof, at a pH of at least about 10, at a temperature in the range from about 20°C to about 45°C, for a period of time from about 30 minutes to about 2 hours, during which reaction the silica gel retains its particulate form, seperating the alkali-reacted, particulate silica g el from the basic solution, and then reacting the said alkali-reacted, silicia gel particles with an aqueous mineral acid solution to remove metal ion from the gel and to activate the gel sufaces with reactive hydroxyl groups.
Claim 8
Dependent Claim
17 words
The process of claim 5 wherein the pH of the strongly basic solution is at least 11.
Claim 9
Dependent Claim
80 words
A process for immobilizing an active enzyme material having available amine groups, at a high density of immobilization per unit volume, comprising reacting a particulate carrier preferably a finely divided silicia gel, that has been activated by the process of claims 1 or 5, with a polyvalent covalent coupling agent to immobilize the coupliny agent, to the activated carrier, then reacting the active enzyme material with the inmbilized coupling agent, to immobilize the active enzyme material to the carrier particles.
Claim 10
Dependent Claim
21 words
The process of claim 7 wherein the coupling agent is selected from the group consisting of an aminosilane and cyanogen bromide.
Claim 11
Dependent Claim
22 words
The process of claim 8, wherein the coupling agent is an aminosilane and the aminosilane is gammaaminopropyltriethoxysilane, and the enzyme is lactase.
Claim 12
Dependent Claim
135 words
In a composite wherein a biologically active material having available amine groups, especially a proteinaceous material, is immobilized to a siliceous inorganic carrier material by covalent bonding to groups pendant from the surface ot the carrier material, the improvement wherein the carrier is a finely divided particulate siliceous inorganic carrier activated by reaction with a strongly basic solution having a pH of at least 10 at an elevated temperature for a sufficient time to generate surface groups of the type SiO - M + , where M is a metallic ion, an ammonium ion, or represents a mixture thereof, in which reaction said carrier retains its particulate form, followed by reaction with a strongly acidic solution to replace the M with hydrogen, so that the carrier particles have available, reactive hydroxyl groups on their sufaces.
Claim 13
Dependent Claim
135 words
In a composite wherein a biologically active material having available amine groups, especially a proteinaceous material, is immobilized to a siliceous inorganic carrier material by covalent bonding to groups pendant from the surface ot the carrier material, the improvement wherein the carrier is a finely divided particulate siliceous inorganic carrier activated by reaction with a strongly basic solution having a pH of at least 10 at an elevated temperature for a sufficient time to generate surface groups of the type SiO - M + , where M is a metallic ion, an ammonium ion, or represents a mixture thereof, in which reaction said carrier retains its particulate form, followed by reaction with a strongly acidic solution to replace the M with hydrogen, so that the carrier particles have available, reactive hydroxyl groups on their sufaces.
Claim 14
Dependent Claim
78 words
The composite of claim 10, wherein the siliceous inorganic carrier material is a silica gel, the carrier has been activated by an initial step of reaction with an aqueous solution of an alkali metal hydroxide at a pH of at least 11 at a temperature in the range from about 20°C to about 45°C for a period of time in the range from about 30 minutes to about 2 hours, and the biologically active material is an enzyme.
Claim 15
Dependent Claim
13 words
The composite. of claim 10, wherein the said proteinaceous material are immunologically reactive.
Claim 16
Dependent Claim
16 words
The composite of claim 10, wherein at least one of said proteinaceous materials is a hormone.
Claim 17
Dependent Claim
20 words
The composite of claim 10, wherein the bonding is the reaction product of an aldehyde-amine reaction and/or an imidoester-amine reaction.
Claim 18
Dependent Claim
67 words
In an immobilized enzyme composite comprising a carrier of solid, free-flowing particles, a first amount of enzyme immobilized to the carrier, and a second amount of enzyme immobilized by covalent coupling through a divalent coupling agent to said first-named enzyme, substantially all of said enzyme retaining characteristic activity, the improvement wherein said first amount of enzyme is immobilized to the carrier by the process of claim 8.
Claim 19
Dependent Claim
19 words
The composite of claim 15, wherein said first-named enzyme and said second-named enzyme are the same enzyme, especially lactase.
Claim 20
Dependent Claim
40 words
The composite of claim 15, wherein there are a plurality of added amounts of enzyme, each added amount being of the same enzyme, and each amount of enzyme, as added, being covalently bonded to the previously added amount of enzyme.
Claim 21
Dependent Claim
9 words
The composite of claim 17,.wherein the enzyme is lactase.
References Cited (Backward Citations)
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