Electrode and method for the detection of an affinity reaction

• US 5,534,132 A
• Filed: 05/04/1995
• Issued: 07/09/1996
• Est. Priority Date: 05/04/1995
• Status: Expired due to Fees

• Current Assignee: Mark Vreeke, Patrick Rocca
• Original Assignee: Mark Vreeke, Patrick Rocca
• Inventors: Rocca, Patrick, Vreeke, Mark
• Primary Examiner(s): Bell, Bruce F.
• Application Number:
  US08/434,782
• Time in Patent Office: 432 Days
• Field of Search: 205/777.5, 204/403, 204/418, 435/817, 435/288, 435/291, 435/4, 435/7.1, 435/7.5, 435/25
• US Class Current: 205/777.5
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

An amperometric biosensor for the detection of an affinity reaction is described. The sensor includes an electrode having on its testing surface a hydrogel in which a selective binding unit and redox species are immobilized.

Claims

1. An electrode for the detection or quantification of a Selective Binding reaction comprising:

an electrode having a testing surface substantially covered with a transducing film comprising an immobilized polymer network which provides for electrical conduction in the absence of diffusional mediators and a selective binding unit (SBU) where the SBU where the SBU has a specific affinity for a chemical, biological agent, or a class of chemicals, and includes immobilizing and oxidoreductase to the SBU'"'"'s conjugate to bind with the SBU, wherein the oxidoreductase is reversibly immobilized at the electrode and electrical conduction between the electrode and the immobilized oxidoreductase occurs.

2. The electrode of claim 1, wherein the immobilizing polymer is a redox polymer containing osmium based redox centers.

3. The electrode of claim 1, wherein the immobilizing polymer is poly(vinyl pyridine) complexed with osmium bis (2,2'"'"'-bipyridine) dichloride.

4. The electrode of claim 1, wherein the immobilizing polymer is poly (N-vinyl imidazole) complexed with osmium bis(2,2'"'"'-bipyridine) dichloride.

5. The electrode of claim 1 where the selective binding unlit is Avidin or streptavidin.

6. The electrode of claim 1 where the selective binding unit is an antibody.

7. The electrode of claim 1 where the selective binding unit is a lectin, specific sugar residue, or single strand of DNA.

8. A method for sensing the occurrence of a binding reaction between a selective binding unit (SBU) and a conjugate to the SBU where changing of the incubation solution is not necessary to differentiate bound and unbound conjugate comprising the steps of coating a current collecting electrode with a hydrogel, the polymer network of which comprises SBUs and multiple reducible and oxidizable centers chemically bound to the network, immersing the hydrogel coated electrode in a solution containing the conjugate, to which an oxidoreductase is covalently bound, allowing the SBU to bind with its conjugate, and measuring the current frown the electrode when the electrolytic solution contains a substrate of the oxidoreductases.

9. A method as in claim 8 where the oxidoreductase label is a peroxidase.

10. A method as in claim 8 where the selective binding occurs between an antigen and an antibody, DNA complimentary strands, or lectin and specific sugar residues.

11. A method as in claim 8 where the selective binding occurs between biotin and avidin.

12. A method as in claim 8 where the selective binding occurs between biotin and an antibody to biotin.

13. A method as in claim 8 where an analyte is detected or quantifies by the analytes competition with a labeled complement for limited binding sites at the affinity electrode.

14. A method as in claim 8 where an analyte is detected or quantified by the analytes binding oxidoreductase labeled complement thus preventing the labeled complement from binding with selective binding units immobilized at the sensor surface.

15. A method as in claim 8 where the modified electrode uses an osmium redox polymer to supply the oxidizable/reducible species.