Enzyme immobilization with a thermochemical-photochemical bifunctional agent

Claim 2

A process for immobilization of an active enzyme on a solid support material which comprises: a. thermochemically attaching to a solid support material an aryl azide bifunctional agent initially possessing both a thermochemically active functional substituent and a photochemically active functional substituent; b. photochemically activating the azide substituent of the product of step (a); and c. reacting the photoactivated azide substituent of the product of step (b) with an enzyme to form a covalent linkage therewith.

Claim 3

The process of claim 1 wherein step (a) is carried out in the absence of visible light of such intensity as to substantially affect the photochemically active functional substituent of the bifunctional agent.

Claim 4

A process in accordance with claim 1 wherein the solid support material is selected from the group consisting of alkylamine glass and aminoethyl cellulose.

Claim 5

A process in accordance with claim 1 wherein the solid support material is alkylamine glass and the bifunctional agent is 1-fluoro-2-nitro-4-azidobenzene.

Claim 6

A process in accordance with claim 1 wherein the solid support material is alkylamine glass and the bifunctional agent is 1-fluoro-2,4-dinitro-5-azidobenzene.

Claim 7

A process in accordance with claim 1 wherein the solid support material is aminoethyl cellulose and the bifunctional agent is 1-fluoro-2-nitro-4-azidobenzene.

Claim 8

A process in accordance with claim 1 wherein the solid support material is aminoethyl cellulose and the bifunctional agent is 1-fluoro-2,4-dinitro-5-azidobenzene.

Claim 9

A process in accordance with claim 1 wherein the solid support material is an agarose.

Claim 10

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 1.

Claim 11

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 3.

Claim 12

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 4.

Claim 13

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 5.

Claim 14

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 6.

Claim 15

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 7.

Claim 16

An active immobilized enzyme in accordance with claim 9 wherein the covalently bound enzyme is asparaginase.

Claim 17

An active immobilized enzyme in accordance with claim 9 wherein the covalently bound enzyme is invertase.

Claim 18

An active immobilized enzyme in accordance with claim 9 wherein the covalently bound enzyme is peroxidase.

Claim 19

An active immobilized enzyme in accordance with claim 9 wherein the covalently bound enzyme is glucose oxidase.

Claim 20

A process for immobilization of an active enzyme on a solid support material which comprises a. reacting alkylamine glass with 1-fluoro-2,4-dinitrobenzene to form a dinitrophenyl derivative of the alkylamine glass; b. reducing the dinitrophenyl derivative to form an aminonitrophenyl derivative of alkylamine glass; c. treating the aminonitrophenyl derivative of alkylamine glass with nitrous acid to form a nitrophenyldiazonium derivative of alkylamine glass; d. substituting the nitrophenyldiazonium derivative with an azide ion to form a nitroazidophenyl derivative of alkylamine glass; e. photochemically activating the nitroazidophenyl derivative of alkylamine glass; and f. reacting the photochemically activated product of step (e) with an enzyme to form a covalent linkage therewith.

Claim 21

A process for immobilization of an active enzyme on a solid support material which comprises a. reacting alkylamine glass with 1-fluoro-2,4-dinitrobenzene to form a dinitrophenyl derivative of the alkylamine glass; b. reducing the dinitrophenyl derivative to form an aminonitrophenyl derivative of alkylamine glass; c. treating the aminonitrophenyl derivative of alkylamine glass with nitrous acid to form a nitrophenyldiazonium derivative of alkylamine glass; d. substituting the nitrophenyldiazonium derivative with an azide ion to form a nitroazidophenyl derivative of alkylamine glass; e. photochemically activating the nitroazidophenyl derivative of alkylamine glass; and f. reacting the photochemically activated product of step (e) with an enzyme to form a covalent linkage therewith.

Claim 22

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 19.

Claim 23

A process for immobilization of an active enzyme on a solid support material which comprises a. reacting aminoethyl cellulose with 1-fluoro-2,4-dinitrobenzene to form a dinitrophenyl derivative of the aminoethyl cellulose; b. reducing the dinitrophenyl derivative to form an aminonitrophenyl derivative of aminoethyl cellulose; c. treating the aminonitrophenyl derivative of aminoethyl cellulose with nitrous acid to form a nitrophenyldiazonium derivative of aminoethyl cellulose; d. substituting the nitrophenyldiazonium derivative with an azide ion to form a nitroazidophenyl derivative of aminoethyl cellulose; e. photochemically activating the nitroazidophenyl derivative of aminoethyl cellulose; and f. reacting the photochemically activated product of step (e) with an enzyme to form a covalent linkage therewith.

Claim 24

A process for immobilization of an active enzyme on a solid support material which comprises a. reacting aminoethyl cellulose with 1-fluoro-2,4-dinitrobenzene to form a dinitrophenyl derivative of the aminoethyl cellulose; b. reducing the dinitrophenyl derivative to form an aminonitrophenyl derivative of aminoethyl cellulose; c. treating the aminonitrophenyl derivative of aminoethyl cellulose with nitrous acid to form a nitrophenyldiazonium derivative of aminoethyl cellulose; d. substituting the nitrophenyldiazonium derivative with an azide ion to form a nitroazidophenyl derivative of aminoethyl cellulose; e. photochemically activating the nitroazidophenyl derivative of aminoethyl cellulose; and f. reacting the photochemically activated product of step (e) with an enzyme to form a covalent linkage therewith.

Claim 25

An active enzyme immobilized on a solid support material produced in accordance with the process of claim 21.