Use of plasma to immobilize protein on polymeric surfaces

• US 5,028,657 A
• Filed: 07/18/1988
• Issued: 07/02/1991
• Est. Priority Date: 07/18/1988
• Status: Expired due to Term

• Current Assignee: BASF AG (BASF SE)
• Original Assignee: ADVANCED RESEARCH & TECHNOLOGY INSTITUTE
• Inventors: Sheu, Min-Shyan, Hsu, Tien-Tsai, Hsiung, Kuang-Pin, Hsiue, Ging H., Wang, Mann-Tchao
• Primary Examiner(s): Nutter, Nathan M.
• Application Number:
  US07/220,570
• Time in Patent Office: 1,079 Days
• Field of Search: 525/54.1, 435/180, 435/181, 435/182, 435/188, 530/402, 436/523, 436/528, 436/529, 436/530, 436/531
• US Class Current: 525/54.1
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: G01N 33/545 Synthetic resin

Abstract

The present invention relates to a method of immobilizing proteins on a polymeric matrix by means of plasma activation and an apparatus and process for the use of such material. The protein mixture is applied to the surface of the polymeric matrix with or without the addition of a crosslinking agent. If is then placed into a plasma generator, wherein the functional groups on both the protein and the matrix molecules are activated to form free radicals. Upon returning from their high energy state, the free radicals form covalent bonds between the proteins and between the protein and the polymeric matrix. Using this method, the proteins are nonspecifically immobilized on the surface of the polymeric matrix. The method can be utilized to immobilize proteins on the surface of polymeric membranes, polymeric beads, polymeric tubes and polymeric plates. The immobilized protein has high biological activity and stability.

Claims

1. A method for the immobilization of a protein on a polymer surface which comprises:

coating said polymer surface with a solution of said protein;

exposing said coated polymer to an environment containing a gas plasma to form free radicals; and

covalently bonding said protein to said polymeric surface at the free radical sites.

2. The method of claim 1 wherein said protein solution is prepared by dissolving the protein in a buffer solution with or without a crosslinking agent.

3. The method of claim 2 wherein said protein solution contains a crosslinking agent.

4. The method of claim 1 wherein said gas plasma is a nitrogen, an oxygen, or an ammonia plasma.

5. The method of claim 1 wherein the temperature of the environment is from 0°

to 25°

C.

6. The method of claim 1 wherein the pressure of the environment is from 0.1 to 0.5 torr.

7. The method of claim 1 wherein the gas plasma is formed in a plasma generator at a power of from 50 to 300 watts.

8. The method of claim 1 wherein said polymer is a polyolefin.

9. The method of claim 1 wherein said polymer is a hydrophobic or hydrophilic polyethylene, polypropylene, or cellulose acetate.

10. The method of claim 1 wherein said polymer is in the form of a membrane, beads, a plate, or a tube.

11. The method of claim 1 wherein the protein is glucose oxidase, lactate oxidase, 1-glutamate decarboxylase, 1-lysine decarboxylase and invertase, mutarotase, or glucose oxidase mixtures.