Method for stabilizing an immobilized fibrinolytic enzyme

• US 4,764,466 A
• Filed: 11/18/1985
• Issued: 08/16/1988
• Est. Priority Date: 11/16/1984
• Status: Expired due to Fees

• Current Assignee: Unitika Limited
• Original Assignee: Unitika Limited
• Inventors: Yabushita, Yasunori, Suyama, Katsuhiko, Takagi, Kunihiko, Koyama, Masanao
• Primary Examiner(s): Naff, David M.
• Application Number:
  US06/799,239
• Time in Patent Office: 1,002 Days
• Field of Search: 435/174, 435/176, 435/177, 435/178, 435/179, 435/180, 435/182, 435/188
• US Class Current: 435/174
• CPC Class Codes: A61K 38/46 Hydrolases (3)
• CPC Class Codes: A61L 2/0011 using physical methods
• CPC Class Codes: A61L 2/0094 Gaseous substances
• CPC Class Codes: C12N 11/00 Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates

Abstract

A fibrinolytic enzyme immobilized on a carrier is stabilized by treating the surface of the carrier on which the enzyme is immobilized with a basic amino acid. The amino acid is preferably histidine, arginine or lysine. Carriers may be inorganic materials, natural polymeric materials or synthetic polymeric materials, in a variety of forms. Fibrinolytic enzymes immobilized include plasmin, brinase, urokinase, streptokinase and tissue plasminogen activator. The stabilized immobilized fibrinolytic enzyme maintains activity better during storage or sterilization.

Claims

1. A method for stabilizing an immobilized fibrinolytic enzyme, comprising contacting the surface of a carrier containing said enzyme immobilized thereon, with a basic amino acid selected from the group consisting of histidine, arginine and lysine.

2. A method according to claim 1, wherein said basic amino acid is histidine.

3. A method according to claim 1, wherein said basic amino acid is arginine.

4. A method according to claim 1, wherein the surface of the carrier is contacted with a solution containing basic amino acid in a range of from about 0.00001 wt% to about 30 wt%.

5. A method according to claim 2, wherein the surface of the carrier is contacted with a solution containing histidine in a range of from about 0.00001 wt% to about 30 wt%.

6. A method according to claim 3, wherein the surface of the carrier is contacted with a solution containing arginine in a range of from about 0.00001 wt% to about 30 wt%.

7. A method according to claim 1, wherein the surface of the carrier is contacted with a solution containing basic amino acid in a range of from 0.001 wt% to 1 wt%.

8. A method according to claim 2, wherein the surface of the carrier is contacted with a solution containing histidine in a range of from 0.001 wt% to 1 wt%.

9. A method according to claim 3, wherein the surface of the carrier is contacted with a solution containing arginine in a range of from 0.001 wt% to 1 wt%.

10. A method according to claim 1, wherein the contacting is carried out at a temperature of from about 0°

to about 50°

C.

11. A method according to claim 4, wherein the contacting is carried out at a temperature of from about 0°

to about 50°

C.

12. A method according to claim 11, wherein said basic amino acid is histidine.

13. A method according to claim 11, wherein said basic amino acid is arginine.

14. A method according to claim 7, wherein the contacting is carried out at a temperature of from about 0°

to about 50°

C.

15. A method according to claim 14, wherein said basic amino acid is histidine.

16. A method according to claim 14, wherein said basic amino acid is arginine.