Novel protein-immobilizing hydrophobic polymeric membrane, process for producing same and apparatus employing same

• US 3,966,580 A
• Filed: 09/16/1974
• Issued: 06/29/1976
• Est. Priority Date: 09/16/1974
• Status: Expired due to Term

• Current Assignee: University of Utah Research Foundation (State of Utah)
• Original Assignee: University of Utah (State of Utah)
• Inventors: Janata, Jiri A., Janata, Jarmila
• Primary Examiner(s): Kaplan, G. L.
• Application Number:
  US05/506,464
• Time in Patent Office: 652 Days
• Field of Search: 204/296, 204/195 M, 204/195 B, 204/1 T, 260/6, 260/8, 117/138.8 B, 195/63, 195/68, 195/DIG. 11, 195/103.5 R-127, 427/322, 427/307, 427/399, 428/474
• US Class Current: 204/403.07
• CPC Class Codes: B01D 67/00931 by introduction of specific groups after membrane formation, e.g. by grafting
• CPC Class Codes: B01D 71/82 characterised by the presence of specified groups, e.g. introduced by chemical after-treatment
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12Q 1/002 Electrode membranes
• CPC Class Codes: C12Q 1/003 Functionalisation
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode
• CPC Class Codes: Y10T 428/31768 Natural source-type polyamide [e.g., casein, gelatin, etc.]

Abstract

A protein-immobilizing hydrophobic polymeric membrane comprising an organic, hydrophobic polymeric substrate with a specific protein reactive group pendant therefrom in preselected concentration is disclosed. The protein reactive groups is a free group of a hydrocarbon chain which has one end thereof absorbed into the surface of said polymeric substrate; said free group, generally at the end of said chain, covalently bonding with a protein molecule. The concentration of protein reactive sites is sufficiently low that the polymeric substrate retains its hydrophobic character. An electrode coated with a protein-immobilized membrane, i.e., a membrane having a hydrocarbon chain to which a protein molecule is attached, provides, in conjunction with a reference electrode, an electrically sensitive system for measuring change in concentration of a compound in solution where said compound selectively reacts with the immobilized protein.

Claims

1. A hydrophobic membrane having one member of an immunochemically reactive pair immobilized thereon comprising:

a. a hydrophobic polymeric substrate swellable by an organic solvent,

b. hydrocarbon chains of at least 6 carbon atoms in length partially absorbed into the surface of said polymeric substrate, said hydrocarbon chains having a reactive site reactive with one compound of an immunochemically reactive pair attached to its non-absorbed portion, and

c. one member of an immunochemically reactive pair reacted with the reactive site of said hydrocarbon chain.

2. The membrane of claim 1 wherein the reactive sites of said hydrocarbon chains unreacted with one compound of an immunochemically reactive pair contain a reaction-blocking molecule.

3. The membrane of claim 1 wherein said reactive site of the hydrocarbon chain is an oxirane group.

4. The membrane of claim 1 wherein said organic polymeric substrate is swellable by aliphatic solvents.

5. The membrane of claim 1 wherein the said hydrocarbon chain is the reaction product of n-decanol and epichlorohydrin.

6. a hydrophobic membrane of claim 1 wherein the compound of an immunochemically reactive pair is a protein.

7. The hydrophobic membrane of claim 1 wherein the compound of an immunochemically reactive pair is an enzyme.

8. The hydrophobic membrane of claim 1 wherein the compound of an immunochemically reactive pair is a monosaccharide.

9. The hydrophobic membrane of claim 1 wherein the compound of an immunochemically reactive pair is a polysaccharide.

10. A quantitative device measuring electric potential difference for detecting presence of compounds reactive with an immobilized compound of an immunochemically reactive pair by utilization of an immunochemical reaction comprising:

a. a reference electrode;

b. a reactive electrode comprising;

1. an electrical metal conductor;

2. a sheath of hydrophobic polymeric membrane comprising;

a. a hydrophobic polymeric substrate swellable by an organic solvent,

b. hydrocarbon chains of at least 6 carbon atoms in length partially absorbed into the surface of said polymeric substrate, said hydrocarbon chains having a reactive site reactive with one compound of an immunochemically reactive pair attached to its non-absorbed portion,

c. one member of an immunochemically reactive pair reacted with the reactive site of said hydrocarbon chain, and

c. electrical conductor means connecting said reference electrode and measuring electrode through a meter sensitive to very slight changes in electrical potential.