Biologically active membrane material

• US 4,066,512 A
• Filed: 05/10/1976
• Issued: 01/03/1978
• Est. Priority Date: 09/06/1974
• Status: Expired due to Term

• Current Assignee: Millipore Investment Holdings, Ltd.
• Original Assignee: Millipore Corporation
• Inventors: Goldin, Stanley M., Lai, Chung Jung
• Primary Examiner(s): Naff, David M.
• Application Number:
  US05/684,746
• Time in Patent Office: 603 Days
• Field of Search: 195/63, 195/68, 195/DIG. 11, 195/103.5 R, 195/127, 424/94, 424/2, 427/338, 427/414, 427/439, 260/2.5 A, 260/6
• US Class Current: 435/10
• CPC Class Codes: B01D 69/144 containing embedded or bound biomolecules
• CPC Class Codes: B01D 71/14 Esters of organic acids
• CPC Class Codes: C12M 21/18 Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
• CPC Class Codes: C12M 23/34 Internal compartments or partitions
• CPC Class Codes: C12M 25/02 Membranes; Filters filters or filtration in general B01D24/00-B01D41/00
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12N 11/02 Enzymes or microbial cells immobilised on or in an organic carrier
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: Y10S 530/816 Attached to the carrier via a bridging agent

Abstract

A composite membrane structure for immobilizing biologically active materials, such as enzymes, is formed by coating a microporous polymeric membrane with a thin layer of an inert proteinaceous material, such as zein or collagen, so that the resultant coated membrane retains intercommunicating capillary pores that extend through its structure. Immobilization of a biologically active material is carried out by contacting the coated membrane with the biologically active material in solution and drying. Biologically active materials immobilized on the membrane can be used to perform biochemical reactions and are useful in carrying out tests for glucose and uric acid.

Claims

1. A composite membrane structure for immobilizing biologically active proteinaceous materials comprising a thin microporous membrane having capillary pore sizes in the range from about 25 nanometers to 14 micrometers, said membrane being formed from a biologically inert synthetic polymer, said membrane having a thin, water-insoluble coating of an inert proteinaceous material on its surfaces, said coating being formed by (a), soaking the membrane with a solution of the proteinaceous material in an organic solvent, and (b), drying to form a thin layer of said inert proteinaceous material such that the coated membrane retains its intercommunicating capillary pores that extend through its structure, to permit fluid flow therethrough, and that provide a high surface-to-volume ratio, said coated membrane being capable of stably immobilizing a biologically active proteinaceous material thereon with retention of its activity upon contacting the coated membrane with a solution in which the biologically active material is dissolved, and drying.

2. A composite membrane in accordance with claim 1 wherein the inert proteinaceous material forming the coating is a natural protein.

3. A composite membrane in accordance with claim 1 wherein the inert proteinaceous material forming the coating is a synthetic polypeptide.

4. A composite membrane structure in accordance with claim 1 wherein the inert proteinaceous material forming the coating is zein or collagen.

5. A composite membrane in accordance with claim 1 having a biologically active proteinaceous material immobilized thereon.

6. A composite membrane in accordance with claim 4, having an enzyme immobilized thereon.

7. A composite membrane in accordance with claim 4, having an antibody immobilized thereon.

8. A composite membrane in accordance with claim 4, having an antigen immobilized thereon.

9. A method for performing an analysis comprising:

selecting a sample suspected of containing a substance in solution to be analyzed for, contacting the sample with said membrane having a biologically active proteinaceous material immobilized thereon in accordance with claim 5, and

thereafter determining whether a reaction occurs between the immobilized proteinaceous material and said substance, which reaction indicates the presence of said substance.

10. A method in accordance with claim 9 wherein the sample is to be analyzed for a substance in solution that is known to change in optical density upon reaction with said immobilized biologically active proteinaceous material, and wherein the step of determining whether a reaction occurred includes measuring the optical density to determine any change that would indicate whether the sample contained the substance in question.

11. A process in accordance with claim 10 wherein the immobilized biologically active material is an enzyme.

12. A process in accordance with claim 11 wherein the substance that is known to change in optical density upon reaction is uric acid, and the immobilized biologically active proteinaceous material is the enzyme uricase.

13. An analytical device comprising a composite membrane structure, having a biologically active proteinaceous material immobilized thereon in accordance with claim 5 said structure being mounted in a holder, a sample chamber disposed in the holder on one side of the composite membrane structure, and a filtrate chamber disposed on the other side of the composite membrane structure.

14. An analytical device in accordance with claim 13 wherein the holder, the membrane and the coating on the membrane are substantially inert relative to the biologically active proteinaceous material that is immobilized on the composite membrane structure.

15. An analytical device comprising:

a cuvette;

a column segment mounted on the upper end of the cuvette, said column segment and cuvette having a configuration permitting the assembly thereof to be placed in a centrifuge, and

a composite membrane structure having a biologically active proteinaceous material immobilized thereon in accordance with claim 5 disposed across the flow path between the column segment and the cuvette.

16. An analytical device in accordance with claim 15 wherein the coating is formed from zein and the immobilized biologically active material is the enzyme uricase.

17. A method for bringing about a molecular interaction between a first proteinaceous material that is biologically active and a second substance that either binds with or is catalytically changed by the first material, comprising:

immobilizing the first material on a composite membrane structure in accordance with claim 1, and

passing a solution of the second material through the composite membrane structure on which the first material is immobilized, to permit binding or a catalytic change to occur.

18. A method in accordance with claim 17 wherein the immobilized first material is an enzyme.