Electrochemical immunosensor and kit and method for detecting biochemical anylyte using the sensor

• US 20030119208A1
• Filed: 12/02/2002
• Published: 06/26/2003
• Est. Priority Date: 12/04/2001
• Status: Abandoned Application

• Current Assignee: Electronics and Telecommunications Research Institute
• Original Assignee: Electronics and Telecommunications Research Institute
• Inventors: Yang, Haesik, Jun, Chi-Hoon, Kim, Youn Tae, Yoon, Hyun Chul
• Application Number:
  US10/308,435
• Publication Number:
  US 20030119208A1
• Time in Patent Office: Days
• US Class Current: 436/524
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 2333/902 Oxidoreductases (1.)
• CPC Class Codes: G01N 2333/908 acting on hydrogen peroxide as acceptor (1.11)
• CPC Class Codes: G01N 2333/916 acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/581 with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)

Abstract

An electrochemical immunosensor including a biological sensor layer with an antigen or a ligand residue immobilized thereon, and a biochemical analyte detection kit and method for electrochemically signaling a biological reaction occurring in the biological sensor layer are provided. The electrochemical immunosensor includes a substrate, an electrode or an electrode array formed on the substrate, and a biological sensor layer formed on the electrode or the electrode array and including a polymeric dendrimer monolayer with an antigen or a ligand residue immobilized on the surface thereof. The biological sensor layer further includes an adhesive layer for biomolecular immobilization between the electrode and the polymeric dendrimer monolayer.

Claims

1. An electrochemical immunosensor comprising:

a substrate;

an electrode or an electrode array formed on the substrate; and

a biological sensor layer formed on the electrode or the electrode array and including a polymeric dendrimer monolayer with an antigen or a ligand residue immobilized on the surface thereof.

2. The electrochemical immunosensor of claim 1, wherein the substrate is silicon or glass.

3. The electrochemical immunosensor of claim 1, wherein the electrode or the electrode array is formed of gold.

4. The electrochemical immunosensor of claim 1, wherein the antigen is immobilized on the surface of the polymeric dendrimer monolayer.

5. The electrochemical immunosensor of claim 4, wherein the antigen contains succinimide or aldehyde functional group.

6. The electrochemical immunosensor of claim 1, wherein the ligand residue is immobilized on the surface of the polymeric dendrimer monolayer.

7. The electrochemical immunosensor of claim 6, wherein the ligand residue is biotin.

8. The electrochemical immunosensor of claim 1, wherein the biological sensor layer further comprises an adhesive layer for biomolecular immobilization between the electrode and the polymeric dendrimer monolayer.

9. The electrochemical immunosensor of claim 8, wherein the adhesive layer for biomolecular immobilization is formed as a self-assembled monolayer basically including thiol or amine group.

17. A method for detecting and quantifying a biochemical analyte in a liquid sample using the electrochemical immunosensor of claim 1, the method comprising:

reacting the liquid sample with the biological sensor layer;

applying a mixture solution of a precipitation substrate and a labeled catalytic enzyme to the surface of the biological sensor layer to induce a precipitate formation reaction; and

detecting an electrochemical signal from the electrode of the electrochemical immunosensor using a cyclic voltammetric method.

18. The method of claim 17, wherein the precipitation substrate is formed of 4-chloro-1-naphthol.

19. The method of claim 17, wherein the labeled catalytic enzyme is peroxidase, alkaline phosphatase, or glucose oxidase.

20. The method of claim 17, wherein detecting the electrochemical signal comprises measuring a change in the voltage-current waveform or maximum current value obtained from the detected electrochemical signal to measure the attenuation of the electrochemical signal due to a reduction in the effective electrode area of the electrochemical immunosensor.

10. A biochemical analyte detection kit comprising:

an electrochemical immunosensor having a biological sensor layer including a self-assembled monolayer formed on an electrode and a polymeric dendrimer monomer with an antigen or a ligand residue immobilized on the surface thereof;

a buffer solution as a dilution of an antibody or a receptor capable of specifically binding to the antigen or the ligand residue in the polymeric dendrimer monolayer, respectively;

a precipitation substrate; and

a labeled catalytic enzyme capable of binding to an antibody or a receptor to discriminate whether a specific interaction between the antigen and the antibody or between the ligand residue and the receptor has occurred and inducing precipitation from the precipitation substrate.

11. The biochemical analyte detection kit of claim 10, wherein the self-assembled monolayer basically includes thiol or amine group.

12. The biochemical analyte detection kit of claim 10, wherein the antigen is immobilized on the polymeric dendrimer monolayer, and an antibody capable of specifically binding to the antigen is diluted in the buffer solution.

13. The biochemical analyte detection kit of claim 10, wherein the ligand residue is immobilized on the polymeric dendrimer monolayer, and a receptor capable of specifically binding to the ligand residue is diluted in the buffer solution.

14. The biochemical analyte detection kit of claim 13, wherein the ligand residue is biotin, and the receptor is avidin or streptavidin.

15. The biochemical analyte detection kit of claim 10, wherein the precipitation substrate is formed of 4-chloro-1-naphthol.

16. The biochemical analyte detection kit of claim 10, wherein the labeled catalytic enzyme is peroxidase, alkaline phosphatase, or glucose oxidase.