Method for fabricating biosensors using hydrophilic polyurethane

• US 6,509,148 B2
• Filed: 02/10/2000
• Issued: 01/21/2003
• Est. Priority Date: 07/09/1998
• Status: Expired due to Term

• Current Assignee: i-SENS, Inc.
• Original Assignee: i-SENS, Inc.
• Inventors: Nam, Hakhyun, Cha, Geun Sig, Shin, Jae Ho
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Padmanabhan, Kartic
• Application Number:
  US09/485,513
• Publication Number:
  US 20020107227A1
• Time in Patent Office: 1,076 Days
• Field of Search: 204/196.06, 204/400, 204/403, 204/410, 204/415, 204/416, 204/418, 204/421, 204/422, 204/424, 204/426, 204/428, 204/429, 204/431, 324/714, 422/68.1, 422/76, 422/82.01, 422/82.02, 422/82.03, 422/82.11, 422/83, 422/90, 435/7.1, 435/7.4, 435/4, 435/7.92, 435/177, 435/180, 435/181, 435/287.2, 435/287.9, 436/85, 436/518, 436/528, 436/531, 436/532, 522/1
• US Class Current: 435/4
• CPC Class Codes: C12Q 1/002 Electrode membranes

Abstract

Disclosed is a method for fabricating biosensors, using hydrophilic polyurethane. Bio-active reagents, including enzymes, antibodies, antigens, cells and receptors, are mixed with hydrophilic polyurethane and the mixture is directly coated over a signal transducer to form a sensing film which serves as a signal detector. The method using hydrophilic polyurethane allows the simplification of the fabrication of biosensors without conducting complicated chemical reactions and washing steps, such as crosslinking. The bio-active reagent entrapped within the hydrophilic polyurethane film can retains its high activity for an extended period of time and the intrinsic potentiometric response of the underlying ion-selective polymeric membrane is not affected by the bio-active reagent immobilized polyurethane film coated on its sensing surface. Therefore, the biosensors are superior in specificity, selectivity, and stability. Also, the adoption of solid-state type electrodes by virtue of the direct immobilization of the bio-active reagents allows the biosensors to be miniaturized, offering the advantages of the detection of small quantities of samples and mass production for low cost.

Claims

1. A method for fabricating a biosensor composed of a signal detector and a signal transducer, comprising:

a) obtaining a mixture solution by dissolving a mixture consisting essentially of hydrophilic polyurethane synthesized from polyethylene glycol, polypropylene glycol, ethylene glycol and diisocyanate, and a bio-active reagent into a solvent selected from the group consisting of tetrahydrofuran, methanol and mixture thereof; and

b) directly immobilizing the mixture on the signal transducer by coating the mixture solution onto the signal transducer and then drying to form a membrane of the hydrophilic polyurethane having the bio-active reagent incorporated therein, wherein the hydrophilic polyurethane stabilizes the bio-active reagent and the bio-active reagent retains activity.

2. The method as set forth in claim 1, wherein the step of obtaining a mixture solution further includes dissolving polylysine in the solvent.

3. The method as set forth in claim 1, wherein the bio-active reagent is an enzyme.

4. The method as set forth in claim 1, wherein the polyethylene glycol and polypropylene glycol are used in a molar ratio range of from 1:

10 to 10;

1.

5. The method as set forth in claim 1, wherein the signal transducer utilizes potentiometry, voltametry or conductometry.

6. The method as set forth in claim 1, wherein the signal transducer is formed of at least one selected from the group consisting of silicon rubber, polyvinyl chloride and its derivatives, aromatic polyurethane and aliphatic polyurethane.

7. The method as set forth in claim 1, wherein the biosensor further comprises one of a conventional electrode and a solid electrode.

8. The method as set forth in claim 3, wherein the enzyme is selected from the group consisting of urease, adenosine deaminase and glucose oxidase.

9. The method as set forth in claim 2, wherein the bio-active reagent is an enzyme.

10. The method as set forth in claim 2, wherein the polyethylene glycol and polypropylene glycol are used in a molar ratio range of from 1:

10 to 10;

1.

11. The method as set forth in claim 2, wherein the signal transducer utilizes potentiometry, voltametry or conductometry.

12. The method as set forth in claim 2, wherein the signal transducer is formed of at least one selected from the group consisting of silicone rubber, polyvinyl chloride and its derivatives, aromatic polyurethane and aliphatic polyurethane.

13. The method as set forth in claim 2, wherein the biosensor further comprises one of a conventional electrode and a solid electrode.

14. The method as set forth in claim 9, wherein the enzyme is selected from the group consisting of urease, adenosine deaminase and glucose oxidase.