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Last Updated: October 28, 2021

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Claims for Patent: 4,885,207

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Summary for Patent: 4,885,207
Title: Biocompatible protein or ligand immobilization system
Abstract:A support matrix generally useful for the immobilization of biologically active proteins is made by coating with a polymeric alcohol a core support of titania or a carbonaceous pyropolymer deposited on a high surface area refractory inorganic oxide, cross-linking the alcohol, and converting a portion of the hydroxyl moieties to sulfonate esters. Such supports covalently bind enzymes and antibodies via a strong carbon-nitrogen single bond to give, for example, an immobilized antibody system extremely resistant to leaching of the antibody, the cross-linked alcohol, or of metals from the core support.
Inventor(s): Johnson; Susan S. (E. Forest Lake, IL), Goodman; James R. (Chicago, IL)
Assignee: UOP (Des Plaines, IL)
Application Number:07/073,251
Patent Claims:1. A method of making a support matrix for immobilization of biologically active proteins comprising coating with a polymeric alcohol a core support which is titania, titanated alumina, or a carbonaceous pyropolymer possessing recurring units containing at least carbon and hydrogen atoms deposited on a high surface area inorganic oxide, cross-linking the polymeric alcohol by reacting it with an hydroxyl-reactive cross-linking agent which is in a molar excess relative to the molar proportion of said polymeric alcohol, converting a portion of the hydroxyl groups of the cross-linked alcohol to a sulfonate ester and recovering the resulting support matrix.

2. The method of claim 1 where the core support is titania.

3. The method of claim 1 where the core support is a carbonaceous pyropolymer deposited on an inorganic oxide selected from the group consisting of alumina, silica-alumina, silica-zirconia, silica, controlled pore glass, hydroxyapatite, zirconia-titania, and alumina-zirconia.

4. The method of claim 3 where the inorganic oxide is alumina.

5. The method of claim 1 where the polymeric alcohol is selected from the group consisting of polyvinyl alcohol, agarose, and cellulose esters containing free hydroxyl groups.

6. The method of claim 5 where the alcohol is polyvinyl alcohol.

7. The method of claim 5 where the polyvinyl alcohol has a molecular weight in the range between about 2,000 and about 25,000.

8. The method of claim 1 where the core support is coated with at least 3% by weight of polyvinyl alcohol.

9. The method of claim 8 where the core support is coated with from about 7 to about 15 weight percent polyvinyl alcohol.

10. The method of claim 1 where the cross-linking agent is selected from: a diglycidyl ether of formula ##STR3## where x is an integer from 1 to about 10; epichlorohydrin; aliphatic diisocyanates of the formula OCN(CH.sub.2).sub.y NCO where y is an integer from 2 to about 10; benzene diisocyanate; toluene diisocyanate; and divinylsulfone.

11. The method of claim 10 where the cross-linking agent is a diglycidyl ether and x is from 2 through about 6.

12. The method of claim 10 where the cross-linking agent is an aliphatic diisocyanate where y is from 4 through about 8.

13. The method of claim 1 where the sulfonate ester is selected from the group consisting of trifluoroethanesulfonate, trifluoromethanesulfonate, para-toluenesulfonate, para-bromobenzenesulfonate, para-nitrobenzenesulfonate, and 2,4-dinitrobenzenesulfonate.

14. The method of claim 1 further characterized in that the sulfonate ester is formed by the reaction with from about 0.1 to about 5.0 molar proportions, based on the hydroxyl moieties in the polymeric alcohol coating, of an activated sulfonic acid with the cross-linked polymeric alcohol.

15. A support matrix resulting from the method of claim 1.

16. An immobilized biologically active protein system having covalently bound biologically active protein resulting from nucleophilic displacement of sulfonate ester groups of the support matrix of claim 15 by a primary amino or sulfhydryl moiety of said protein.

17. The immobilized biologically active protein system of claim 16 where the protein is an enzyme, an antibody, or an antigen.

18. The immobilized biologically active protein system of claim 17 where the protein is an enzyme selected from the group consisting of glucose isomerase, glucoamylase, lactase, cellulase, glucose oxidase, peroxidase, ribonuclease, urease, histidase, trypsin, papain, hexakinase, chymotrypsin, acylase, invertase, ficin, lysozyme, protease, pepsin, rennin, xylanase, beta amylase, gamma amylase, asparaginase, cholesterol oxidase, alcohol dehydrogenase, amino acid oxidase, collagenase, arginase, catalase, deoxyribonuclease, heparinase, uricase, glutaminase, bilirubin oxidase, B-glucuronidase, phenylalanine ammonia lyase, arginase, UDP glucurolnyl transferase, streptokinase, tyrosinase, alpha-galactosidase, urokinase, superoxide dismutase, fibrinolysin and carboxypeptidase G-1.

19. The immobilized biologically active protein system of claim 17 where the protein is an antibody selected from the group consisting of I.sub.g A, I.sub.g E, I.sub.g M, I.sub.g D and I.sub.g G immunoglobulins and F.sub.ab fragments.

Details for Patent 4,885,207

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Microbix Biosystems Inc. KINLYTIC urokinase For Injection 021846 1978-01-16 ⤷  Free Forever Trial 2039-03-29
Recordati Rare Diseases, Inc. ELSPAR asparaginase For Injection 101063 1978-01-10 ⤷  Free Forever Trial 2039-03-29
Smith & Nephew, Inc. SANTYL collagenase Ointment 101995 1965-06-04 ⤷  Free Forever Trial 2039-03-29
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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