Plasma-induced protein immobilization on polymeric surfaces

• US 5,306,768 A
• Filed: 08/11/1992
• Issued: 04/26/1994
• Est. Priority Date: 07/18/1988
• Status: Expired due to Term

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research 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:
  US07/929,159
• Time in Patent Office: 623 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, 422/50, 422/68.1, 422/70
• US Class Current: 422/50
• 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. It 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 surfaces of polymeric membranes, polymeric beads, polymeric tubes and polymeric plates. The immobilized protein has high biological activity and stability.

Claims

1. An apparatus for treating, sensing, assaying, or transforming a biological substance which comprises (a) a reaction chamber;

(b) means for supporting a protein immobilized on the surface of a polymer in said reaction chamber;

(c) means for introducing said biological substance into said reaction chamber so as to contact said immobilized protein;

said protein immobilized to the surface of the polymer being prepared by coating said polymeric 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 free radical sites.

2. The apparatus of claim 1 wherein said exposure takes place in the presence of a crosslinking agent.

3. The apparatus of claim 1 wherein said plasma is a nitrogen, an oxygen, or an ammonia plasma.

4. The apparatus of claim 1 wherein the exposure temperature is from 0°

to 25°

C.

5. The apparatus of claim 1 wherein the exposure pressure is from 0.1 to 0.2 torr.

6. The apparatus of claim 1 wherein the power generated during said exposure is from 50 to 300 watts.

7. The apparatus of claim 1 wherein the polymer is in the form of a membrane.

8. The apparatus of claim 1 wherein the polymer is a bead.

9. The apparatus of claim 1 wherein the polymer is a plate.

10. The apparatus of claim 1 wherein the polymer is a tube.

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