Immobilization of proteins on polymeric supports
AssigneeAkzona Incorporated
InventorsLincoln D. Metcalfe, Dieter Frank
Life of this patent
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Abstract
A protein is immobilized by a simple and effective method on a porous polymeric support by a first soaking in a dilute long-chain cationic solution and a second soaking in a dilute aqueous protein solution. The long chain cationic is preferably a nitrogen compound such as a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms. A preferred diamine is N-coco-1,3-diamino-propane. The protein may be an enzyme or hormone. A preferred enzyme is catalase or a two enzyme system of catalase and glucose oxidase.
Claims28
A method of immobilizing a protein on a porous polymeric support, comprising a first soaking step in which said support is placed in a dilute solution of a long-chain cationic comprising a nitrogen compound having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent, said support being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a dilute aqueous solution of said protein.
A method of immobilizing a hormone on a support, comprising a first soaking step in which said support is placed in a dilute solution of a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent for said diamine for between 5 and 60 minutes, said diamine solution being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a aqueous solution of said hormone, said aqueous solution having a concentration of said hormone of from 0.002% (w/v) to 0.2% (w/v).
A method of immobilizing an enzyme on a porous polymeric support, comprising a first soaking step in which said support is placed in a dilute solution of a long-chain cationic comprising a nitrogen compound having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent, said support being gently agitated during said first soaking step, and a second soaking step in which said support is soaked in a dilute aqueous solution of said enzyme.
A method of immobilizing catalase on a porous polymeric support, comprising a pretreatment step in which said support is placed in a dilute solution of a metal salt in a salt solvent for between 5 and 10 minutes, rinsing said support with said salt solvent or another salt solvent, a first soaking step in which said support is placed in a dilute solution of a diamine having at least one alkyl or alkenyl group containing at least eight carbon atoms attached thereto in a solvent for said diamine for between 10 and 60 minutes, said diamine solution being gently agitated during said first soaking step, and a second soaking step in which said support is soaked with agitation in a 0.8% (w/v) aqueous catalase solution.
A method of immobilizing catalase on a porous polymeric film support, comprising a pretreatment in which said support is placed in a 1% (w/v) metal salt solution of SnCl.sub.2.2H.sub.2 O in acetone;
- removing said support from said metal salt solution after five minutes and rinsing said support with acetone;
- a first soaking step in which said support is immersed for 60 minutes in a 1% (w/v) diamine solution of N-coco-1,3-diaminopropane in chloroform;
- removing said support from said diamine solution and wetting said support with isopropanol;
- and a second soaking step in which said support is immersed for more than four hours in a 0.8% (v/v) aqueous catalase solution.
A method for immobilizing glucose oxidase and catalase on a porous polymer powder support, comprising:
- a first soaking step in which said support is immersed for 10 minutes in a 1% (w/v) diamine solution of N-coco-1,3-diaminopropane in chloroform;
- removing said support from said diamine solution and wetting said support;
- and a second soaking step in which said support is immersed for more than four hours in a dilute buffer solution of 2.5% (v/v) glucose oxidase and 0.5% (v/v) catalase in deionized water.
Dependencies are inferred from the claim text.