Antibody covalently bound film immunobiosensor

• US 5,567,301 A
• Filed: 03/01/1995
• Issued: 10/22/1996
• Est. Priority Date: 03/01/1995
• Status: Expired due to Term

• Current Assignee: Illinois Institute of Technology
• Original Assignee: Government of the United States of America, Illinois Institute of Technology
• Inventors: Gendel, Steven M., Stetter, Joseph R., Hesketh, Peter J., Maclay, G. Jordan
• Primary Examiner(s): Bell, Bruce F.
• Application Number:
  US08/396,229
• Time in Patent Office: 601 Days
• Field of Search: 204/403, 204/415, 204/153.12, 435/817, 435/288, 435/291, 435/4, 435/7.1, 435/7.93
• US Class Current: 205/777.5
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

A biosensor comprising a substrate material, at least one antibody covalently immobilized on the substrate material, and a pair of metal contact electrodes for measuring the impedance of the biosensor. A process for producing a biosensor in accordance with this invention as well as a process for utilizing the biosensor are also disclosed.

Claims

1. A biosensor comprising:

a substrate material;

at least one covalently bound antibody immobilized on said substrate material; and

impedance detection means for measuring an impedance of said biosensor.

2. A biosensor in accordance with claim 1, wherein said substrate material is a material capable of being processed by at least one semiconductor device process.

3. A biosensor in accordance with claim 2, wherein said substrate material is a planar semiconductor device.

4. A biosensor in accordance with claim 3, wherein said substrate material comprises an insulator layer passivated on a silicon chip.

5. A biosensor in accordance with claim 4, wherein said insulator layer is silicon dioxide.

6. A biosensor in accordance with claim 4 further comprising a discontinuous ultra-thin metal film layer deposited on said insulator layer.

7. A biosensor in accordance with claim 6, wherein said discontinuous ultra-thin metal film layer comprises a plurality of distributed islands of metal with an uncovered portion of said insulator layer disposed between neighboring said islands.

8. A biosensor in accordance with claim 7, wherein a size of said uncovered portion of said insulator layer between said neighboring islands corresponds to a size range of said at least one covalently bound antibody.

9. A biosensor in accordance with claim 6, wherein said metal is selected from the group consisting of platinum, palladium, gold, iridium, rhodium, osmium and mixtures thereof.

10. A biosensor in accordance with claim 6, wherein said at least one covalently bound antibody is disposed on said discontinuous ultra-thin metal film layer.

11. A biosensor in accordance with claim 6, wherein said impedance detection means comprises two metal contact electrodes disposed on said discontinuous ultra-thin metal film layer.

12. A biosensor in accordance with claim 4, wherein said impedance detection means comprises two metal contact electrodes disposed on said insulator at a distance from one another.

13. A biosensor in accordance with claim 12, wherein said at least one covalently bound antibody is disposed on at least one of said insulator layer and said two metal contact electrodes.

14. A process for producing a biosensor for measuring antigen-antibody binding reactions comprising:

thermally growing an insulator layer on a silicon chip substrate;

depositing a discontinuous ultra-thin metal film layer on said insulator;

securing two metal contact-electrodes at a distance from each other on said discontinuous ultra-thin metal film layer; and

covalently immobilizing at least one antibody bioactive layer on said discontinuous ultra-thin metal layer.

15. A process in accordance with claim 14, wherein said insulator layer is a silicon dioxide layer thermally grown in a dry oxygen atmosphere at a temperature in the range of about 900°

C. to 1300°

C.

16. A process in accordance with claim 14, wherein said metal is selected from the group consisting of platinum, palladium, gold, iridium, rhodium, osmium and mixtures thereof.

17. A process in accordance with claim 16, wherein said metal is platinum.

18. A process in accordance with claim 14, wherein said discontinuous ultra-thin metal film layer comprises a plurality of distributed islands of metal with an uncovered portion of said insulator layer disposed between neighboring said islands.

19. A process in accordance with claim 14, wherein said discontinuous ultra-thin metal film layer is electrically deposited on said insulator layer by one of sputtering and evaporation.

20. A process in accordance with claim 14, wherein said at least one antibody is anti-SEB.

21. A process in accordance with claim 20, wherein said anti-SEB layer is immobilized by silanization of the surface of said discontinuous ultra-thin metal film layer followed by reaction of said silanized said discontinuous ultra-thin metal film layer with a crosslinking agent and then said anti-SEB.

22. A process in accordance with claim 21, wherein said anti-SEB is covalently bound to said silanized discontinuous ultra-thin metal film layer by a chemical bond.

23. A process in accordance with claim 22, wherein said anti-SEB is covalently bound to said silanized discontinuous ultra-thin metal film layer by a peptide bond.

24. A process in accordance with claim 14, wherein said two metal contact-electrodes are connected to a header through gold-plated copper wires with silver epoxy and cured at about 150°

C. for about 30 minutes.

25. A process for measuring antigen-antibody binding reactions comprising:

contacting a material comprising a chosen antigen with a biosensor comprising a substrate material, at least one covalently bound antibody corresponding to said chosen antigen immobilized on said substrate material, and impedance detection means for measuring an impedance of said biosensor; and

measuring the impedance between two metal contact-electrodes of said biosensor.