Claims for Patent: 4,098,645

Summary for Patent: 4,098,645

Title:	Immobilization of proteins with polyurethane polymers
Abstract:	A protein which can be an enzyme is immobilized by: (a) admixing the protein and an isocyanate-capped liquid polyurethane prepolymer in the absence of water to form a resulting mixture (an intermediate product); and (b) forming the intermediate product by reacting it with water to form a polyurethane foam comprising the immobilized enzyme. When certain proteins in sufficient amount are mixed with the prepolymer in the absence of water the resultant protein prepolymer mixture will solidify to produce a solid non-foamed product containing a protein immobilized therein. Initially mixing the protein and prepolymer in the absence of water results in immobilization of a substantially greater amount of protein than when water is present.
Inventor(s):	Hartdegen; Frank Joseph (Columbia, MD), Swann; Wayne Elliott (Pasadena, MD)
Assignee:	W. R. Grace & Co. (New York, NY)
Application Number:	05/743,035
Patent Claims:	1. In a process for preparing an immobilized protein comprising admixing an isocyanate-capped liquid polyurethane prepolymer, the protein, and water to produce a polyurethane foam with the protein integrally bound to the polyurethane foam, the improvement comprising: (a) admixing the liquid polyurethane prepolymer and the protein in the absence of water to form a resulting solution; and (b) foaming the resulting solution by admixing it with an amount of water effective for producing foaming. 2. The process of claim 1 in which the immobilized protein is washed to remove unbound protein and to hydrolyze any unreacted isocyanate groups. 3. The process of claim 1 in which the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate and a polyethlene glycol. 4. The process of claim 1 in which the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate and a polyethylene glycol having a molecular weight of about 800-1,200. 5. The process of claim 1 in which the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate and a member selected from the group consisting of a polyoxybutylene polyol polymer, ethylene glycol, diethylene glycol, a polyoxyethylene polyol polymer, pentaerythritol, glycerol, trimethylolpropane, and a polyoxypropylene polyol polymer. 6. The process of claim 1 in which the protein is an enzyme, an antibody, or an antigen. 7. The process of claim 1 in which the protein is human immunoglobulin G. 8. The process of claim 1 in which the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate with an admixture of polyethylene glycol having a molecular weight of about 800-1,200 and trimethylolpropane, the trimethylolpropane and the polyethylene glycol being provided in a mole ratio of about 1:1-4 and the toluene diisocyanate being provided at a rate of about 0.85-1.25 mole per equivalent of --OH provided by the polyethylene glycol plus the trimethylolpropane. 9. The process of claim 1 in which the protein is an enzyme and the enzyme is urease, trypsin, lactase, glucose oxidase, chymotrypsin, ribonuclease, peroxidase, pepsin, rennin, invertase, papain, asparaginase, pectinase, pectin esterase, penicillin amidase, glucose isomerase, lysozyme, amino acid acylase, pronase, alcohol dehydrogenase, .alpha.-amylase, .beta.-amylase, subtilisin, amino acid oxidase, catalase, tannase, phenol oxidase, glucoamylase, pullulanase, cellulase, ficin, bromelain, pancreatin, isoamylase, lipase, malic dehydrogenase, hexokinase, lactate dehydrogenase, adenosine deaminase, uricase, galactose oxidase, diaphorase, cholinesterase, aldolase, pyruvate carboxylase, phospharylase, cephalosporin amidase, isocitric dehydrogenase, .alpha.-glycerolphosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, malic enzyme, glucose-6-phosphate dehydrogenase, 5-dehydroshikimic reductase, glutathione reductase, glycolic acid oxidase, yeast cytochrome c reductase, luciferase, nitrite reductase, glutamyl transferase, glutathione synthetase, glycocyamine phosphokinase, hippuric acid synthetase, aldehyde oxidase, succinic dehydrogenase, nitrate reductase, xanthine oxidase, lipoyl dehydrogenase, flavin peroxidase, glycine oxidase, carboxylase, .alpha.-keto acid dehydrogenase, or transketolase. 10. The process of claim 1 in which the isocyanate-capped liquid polyurethane prepolymer is made by reacting toluene diisocyanate with a polyhydroxy compound selected from the group consisting of polyoxybutylene polyol polymer, ethylene glycol, diethylene glycol, polyoxyethylene polyol polymer, pentaerythritol, glycerol, trimethylol propane and polyoxypropylene polyol polymer. 11. A process for preparing an immobilized protein comprising: (a) forming a first product by admixing, in the absence of water, the protein and a liquid polyisocyanate; (b) forming a second product comprising an isocyanate-capped liquid polyurethane prepolymer with the protein dissolved therein by admixing and reacting, in the absence of water, the first product and an amount of a polyol effective for forming the second product; and (c) forming the immobilized protein by admixing the second product and an amount of water effective for producing foaming. 12. The process of claim 11 in which the protein is an enzyme, an antibody, or an antigen. 13. A process for preparing an immobilized protein comprising: (a) forming a first product by admixing, in the absence of water, the protein and a liquid polyol; (b) forming a second product comprising an isocyanate-capped liquid polyurethane prepolymer with the protein dissolved therein by reacting, in the absence of water, the first product and an amount of a polyisocyanate effective for forming the second product; and (c) forming the immobilized protein by admixing the second product and an amount of water effective for producing foaming. 14. The process of claim 13 in which the protein is an enzyme, an antibody, or an antigen. 15. In a process for preparing an immobilized enzyme comprising admixing an isocyanate-capped liquid polyurethane prepolymer, the enzyme, and water to produce a polyurethane foam with the enzyme integrally bound to the polyurethane foam, the improvement comprising: (a) admixing, in the absence of water, the liquid polyurethane prepolymer and a substrate reactable with the enzyme to form a first composition; (b) admixing, in the absence of water, the first composition and the enzyme to form a second composition; and (c) foaming the second composition by admixing it with an amount of water effective for producing foam to form a poly(urea-urethane) foam comprising the immobilized enzyme. 16. The process of claim 15 in which the enzyme is penicillin amidase and the substrate is benzylpenicillin. 17. The process of claim 15 in which the enzyme is glucoamylase and the substrate is starch. 18. The process of claim 15 in which the enzyme is glucoseisomerase and the substrate is starch. 19. The process of claim 15 in which the enzyme is amino acid acylase and the substrate is an acylated amino acid. 20. A method of preparing biologically active polyurethanes comprising dissolving an effective amount of a biologically active protein in an isocyanate-capped liquid urethane prepolymer in the absence of water, and allowing said protein and prepolymer to react to form a solid non-foamed product, said protein characterized as being both soluble in said prepolymer and reactive therewith to yield a solid polyurethane wherein the protein is bound to the polymer in active form. 21. A method as in claim 20 wherein the protein is penicillin amidase. 22. A method as in claim 20 wherein the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate and a polyethylene glycol. 23. A method as in claim 20 wherein the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate and a member selected from the group consisting of a polyoxybutylene polyol polymer, ethylene glycol, diethylene glycol, a polyoxyethylene polyol polymer, pentaerythritol, glycerol, trimethylolpropane, and a polyoxypropylene polyol polymer. 24. A method as in claim 20 wherein the isocyanate-capped liquid polyurethane prepolymer is prepared by reacting toluene diisocyanate with an admixture of polyethylene glycol having a molecular weight of about 800-1,200 and trimethylolpropane, the trimethylolpropane and the polyethylene glycol being provided in a mole ratio of about 1:1-4 and the toluene diisocyanate being provided at a rate of about 0.85-1.25 mole per equivalent of --OH provided by the polyethylene glycol plus the trimethylolpropane.

Details for Patent 4,098,645

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	ELSPAR	asparaginase	For Injection	101063	01/10/1978		1996-02-24
Csl Behring Ag	CARIMUNE, CARIMUNE NF, PANGLOBULIN, SANDOGLOBULIN	immune globulin intravenous (human)	For Injection	102367	07/27/2000		1996-02-24
Csl Behring Ag	PRIVIGEN	immune globulin intravenous (human), 10% liquid	Injection	125201	07/26/2007		1996-02-24
Csl Behring Ag	PRIVIGEN	immune globulin intravenous (human), 10% liquid	Injection	125201	10/02/2009		1996-02-24
Csl Behring Ag	PRIVIGEN	immune globulin intravenous (human), 10% liquid	Injection	125201	02/07/2013		1996-02-24