US4206286A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Immobilization of proteins on inorganic supports
Assignee: Bayer Corp
•
Filing: 1977-11-14
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Granted: 1980-06-03
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Expires: 1997-11-14
Abstract
Abstract A method of producing an immobilized protein composite, i.e. enzymes such as glucose oxidase, on an alumina support, comprising modifying some of the free functional groups of the protein such as amino groups, by reaction with an acid or anhydride, depositing the modified protein on a porous inorganic support at controlled pH and temperature, and cross-linking remaining free functional groups of the protein with adjacent deposited protein functional groups to form interprotein peptide bonds. The resultant composite has long-term continuous use.
Inventors
- 👤 Melvin H. Keyes
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 12:22
CPC Classifications
C; C12; C12N; C12N11/00; C12N11/14
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
130 words
A method for preparing an immobilized composite from a protein having free amino-functional and carboxyl-functional groups, comprising modifying said protein by reacting at least some of said amino-functional groups with a modifying agent selected from a group consisting of: succinic anhydride, succinic acid, malic acid, and glutaric anhydride, to produce a modified protein while leaving some of said amino-functional groups unmodified, depositing said modified protein on the surface of a porous, inorganic support having a high surface area by maintaining said composite within a pH and temperature range which facilitates deposition of said protein, and directly reacting said unmodified groups of said deposited protein with free functional groups in the presence of a carbodiimide agent to form peptide bonds between said deposited proteins, whereby an immobilized protein-support composite is prepared.
Claim 2
Independent Claim
130 words
A method for preparing an immobilized composite from a protein having free amino-functional and carboxyl-functional groups, comprising modifying said protein by reacting at least some of said amino-functional groups with a modifying agent selected from a group consisting of: succinic anhydride, succinic acid, malic acid, and glutaric anhydride, to produce a modified protein while leaving some of said amino-functional groups unmodified, depositing said modified protein on the surface of a porous, inorganic support having a high surface area by maintaining said composite within a pH and temperature range which facilitates deposition of said protein, and directly reacting said unmodified groups of said deposited protein with free functional groups in the presence of a carbodiimide agent to form peptide bonds between said deposited proteins, whereby an immobilized protein-support composite is prepared.
Claim 3
Dependent Claim
21 words
The method according to claim 1, wherein said support is -60+70 mesh, U.S. Standard Sieve, having about 0.1 micron diameter pores.
Claim 4
Dependent Claim
23 words
The method according to claim 1, wherein said support is -60+70 mesh, U.S. Standard Sieve, porous alumina having about 0.1 micron diameter pores.
Claim 5
Dependent Claim
12 words
The method according to claim 1, wherein said carbodiimide is 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide HCl.
Claim 6
Dependent Claim
12 words
The method according to claim 1, wherein said protein is an enzyme.
Claim 7
Dependent Claim
12 words
The method according to claim 1, wherein said protein is glucose oxidase.
Claim 8
Dependent Claim
12 words
The method according to claim 1, wherein said inorganic support is alumina.
Claim 9
Dependent Claim
30 words
The method according to claim 7, wherein said alumina is activated prior to the deposition of said protein by contact with aqueous hydrochloric acid for a sufficient period of time.
Claim 10
Dependent Claim
16 words
The method according to claim 1, wherein said pH range is about 3.5 to about 9.
Claim 11
Dependent Claim
19 words
The method according to claim 1, wherein said pH is in the range of about 5 to about 8.
Claim 12
Dependent Claim
18 words
The method according to claim 1, wherein said temperature is in the range between 5° and 50° C.
Claim 13
Dependent Claim
18 words
The method according to claim 1, wherein said temperature is in the range between 15° and 25° C.
Claim 14
Dependent Claim
13 words
The method according to claim 1, wherein said carbodiimide is 1-cyclohexyl-3-(2-murpholinethyl)-carbodiimide metho-p-toluene sulfonate.
Claim 15
Dependent Claim
13 words
An immobilized protein composite made in accordance with the method of claim 1.
Claim 16
Dependent Claim
12 words
The immobilized composite of claim 14, wherein said protein is an enzyme.
Claim 17
Dependent Claim
12 words
The immobilized composite of claim 14, wherein said protein is glucose oxidase.
Claim 18
Dependent Claim
11 words
An immobilized composite of claim 14, wherein said support is alumina.
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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) | 1983-06-03 | 1983-12-03 | 1984-06-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-06-03 | 1987-12-03 | 1988-06-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-06-03 | 1991-12-03 | 1992-06-03 | $7700.00 | Window Closed / Lapsed |