Biosensor for detecting the presence of chosen analytes

• US 5,736,342 A
• Filed: 11/08/1995
• Issued: 04/07/1998
• Est. Priority Date: 09/21/1993
• Status: Expired due to Fees

• Current Assignee: Washington State University
• Original Assignee: Washington State University
• Inventors: Davis, William C., Sutisna, Himawan, Koch, Alan R., Reiken, Steven R., Moffett, David F., Van Wie, Bernard J., Silber, Moris
• Primary Examiner(s): Allen, Marianne P.
• Assistant Examiner(s): DUFFY, PATRICIA ANN
• Application Number:
  US08/555,347
• Time in Patent Office: 881 Days
• Field of Search: 204/403-418, 356/318, 435/7.21, 435/7.9-7.95, 435/817, 435/7.2, 435/7.8, 436/518, 436/528, 436/532, 436/533, 436/806, 436/501, 436/512, 436/519
• US Class Current: 435/7.2
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/944 Acetylcholine
• CPC Class Codes: Y10S 435/819 Fermentation vessels in series
• CPC Class Codes: Y10S 436/819 Multifunctional antigen or antibody

Abstract

The present invention relates to a biosensor for detecting the presence of an analyte in solution. The biosensor is composed of:

a) an ion channel receptor immobilized in a suspended lipid film which is in contact with an ion-containing solution on each side thereof;

b) a ligand, conjugated to the receptor, which has particular affinity for the analyte in solution, and, which, upon binding to the analyte in solution, causes an inhibition in ion flow through the channel; and

c) means to detect the inhibition of ion flow through the channel in response to the binding of conjugated ligand and analyte.

Claims

1. A biosensor for use in a non-competitive immunoassay for detecting the presence of a chosen analyte in solution which comprises:

a) an ion channel receptor immobilized in a suspended lipid film which film is in contact with an ion-containing solution on each side thereof;

b) a ligand, conjugated to the receptor, said ligand which specifically binds to the analyte in solution, and, which, upon binding to the analyte in solution, causes an inhibition in ion flow through the ion channel; and

c) means to detect the inhibition of ion flow through the ion channel in response to the binding of conjugated ligand and analyte.

2. A biosensor as claimed in claim 1 wherein the receptor is the nicotinic acetylcholine receptor.

3. A biosensor as claimed in claim 1 wherein the lipid film is in the form of a membrane.

4. A biosensor as claimed in claim 2 wherein the lipid film is in the form of a membrane.

5. The biosensor as claimed in claim 1 wherein the ligand which is conjugated to the receptor is a bispecific antibody which has one arm which specifically binds the receptor and another arm which specifically bids the chosen analyte to be detected in solution.

6. The biosensor as claimed in claim 2 wherein the ligand which is conjugated to the receptor is a bispecific antibody which has one arm which specifically binds the receptor and another arm which specifically binds the chosen analyte to be detected in solution.

7. The biosensor as claimed in claim 3 wherein the ligand which is conjugated to the receptor is a bispecific antibody which has one arm which specifically binds the receptor and another arm which specifically binds the chosen analyte to be detected in solution.

8. The biosensor as claimed in claim 4 wherein the ligand which is conjugated to the receptor is a bispecific antibody which has one arm which specifically binds the receptor and another arm which specifically binds the chosen analyte to be detected in solution.

9. The biosensor as claimed in claim 1 wherein the ligand which is conjugated to the receptor is a Fab'"'"' fragment which specifically binds with the chosen analyte to be detected in solution.

10. The biosensor as claimed in claim 2 wherein the ligand which is conjugated to the receptor is a Fab'"'"' fragment which specifically binds with the chosen analyte to be detected in solution.

11. The biosensor as claimed in claim 3 wherein the ligand which is conjugated to the receptor is a Fab'"'"' fragment which specifically binds with the chosen analyte to be detected in solution.

12. The biosensor as claimed in claim 4 wherein the ligand which is conjugated to the receptor is a Fab'"'"' fragment which specifically binds with the chosen analyte to be detected in solution.

13. A biosensor as claimed in claim 1 wherein the detection means (c) are electronic.

14. A biosensor as claimed in claim 1 wherein the detection means (c) are optical.

15. The biosensor as claimed in claim 1 wherein the ligand, which is conjugated to the receptor, is a Fab'"'"' fragment and the Fab'"'"' fragment is also linked to a hapten to which the chosen analyte to be detected in solution will bind.

16. The biosensor as claimed in claim 2 wherein the ligand, which is conjugated to the receptor, is a Fab'"'"' fragment and the Fab'"'"' fragment is also linked to a hapten to which the chosen analyte to be detected in solution will bind.

17. The biosensor as claimed in claim 3 wherein the ligand, which is conjugated to the receptor, is a Fab'"'"' fragment and the Fab'"'"' fragment is also linked to a hapten to which the chosen analyte to be detected in solution will bind.

18. The biosensor as claimed in claim 4 wherein the ligand, which is conjugated to the receptor, is a Fab'"'"' fragment and the Fab'"'"' fragment is also linked to a hapten to which the chosen analyte to be detected in solution will bind.