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Construction method of nanoprobe C60 based electrochemical immunosensor for carcino-embryonic antigens

A technology of immunosensor and nanoprobe, which is applied in the field of electrochemical immunosensor construction, can solve the problems of complex operation, high price, and unsuitability for real-time detection, and achieve the effects of increasing sensitivity, avoiding leakage, and promoting electron transfer

Inactive Publication Date: 2016-06-15
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional immunoassay methods usually have complex operations, are expensive, and are not suitable for real-time detection

Method used

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  • Construction method of nanoprobe C60 based electrochemical immunosensor for carcino-embryonic antigens
  • Construction method of nanoprobe C60 based electrochemical immunosensor for carcino-embryonic antigens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 A kind of based on nanoprobe C 60 Construction method of the CEA electrochemical immunosensor

[0032] (1) with Al 2 o 3 Grind a glassy carbon electrode (GCE) with a diameter of 4 mm with polishing powder, ultrasonically clean it in ethanol and ultrapure water for 3 min, and dry it with nitrogen; use 10 mL of HAuCl with a concentration of 1.0 mmoL / L 4The solution was used as the bottom solution, GCE was used as the working electrode, the Ag / AgCl electrode was used as the reference electrode, and the platinum electrode was used as the counter electrode. After 30 circles of cyclic voltammetry scanning in the potential range of -0.8 ~ 0.6V, the electrode was placed at 0.1moL / L Soak in KCl solution overnight, wash with PBS (pH7.4) buffer solution to obtain gold nanocluster modified glassy carbon electrode (nano-Au / GCE);

[0033] (2) Take 10 μL of 1.0 μg / mL CEA primary antibody (Ab 1 ) standard solution onto the surface of nano-Au / GCE, incubated in a refrige...

Embodiment 2

[0037] Embodiment 2 A kind of based on nanoprobe C 60 Construction method of the CEA electrochemical immunosensor

[0038] (1) with Al 2 o 3 Grind a glassy carbon electrode (GCE) with a diameter of 4 mm with polishing powder, ultrasonically clean it in ethanol and ultrapure water for 3 min, and dry it with nitrogen; use 10 mL of HAuCl with a concentration of 1.0 mmoL / L 4 The solution was used as the bottom solution, GCE was used as the working electrode, the Ag / AgCl electrode was used as the reference electrode, and the platinum electrode was used as the counter electrode. After 30 circles of cyclic voltammetry scanning in the potential range of -0.8 ~ 0.6V, the electrode was placed at 0.1moL / L Soak in KCl solution overnight, wash with PBS (pH7.4) buffer solution to obtain gold nanocluster modified glassy carbon electrode (nano-Au / GCE);

[0039] (2) Take 10 μL of 2.0 μg / mL CEA primary antibody (Ab 1 ) standard solution onto the surface of nano-Au / GCE, incubated in a refrig...

Embodiment 3

[0043] Embodiment 3 A kind of based on nanoprobe C 60 Construction method of the CEA electrochemical immunosensor

[0044] (1) with Al 2 o 3 Grind a glassy carbon electrode (GCE) with a diameter of 4 mm with polishing powder, ultrasonically clean it in ethanol and ultrapure water for 3 min, and dry it with nitrogen; use 10 mL of HAuCl with a concentration of 1.0 mmoL / L 4 The solution was used as the bottom solution, GCE was used as the working electrode, the Ag / AgCl electrode was used as the reference electrode, and the platinum electrode was used as the counter electrode. After 30 circles of cyclic voltammetry scanning in the potential range of -0.8 ~ 0.6V, the electrode was placed at 0.1moL / L Soak in KCl solution overnight, wash with PBS (pH7.4) buffer solution to obtain gold nanocluster modified glassy carbon electrode (nano-Au / GCE);

[0045] (2) Take 10 μL of 2.5 μg / mL CEA primary antibody (Ab 1 ) standard solution onto the surface of nano-Au / GCE, incubated in a refrig...

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Abstract

The invention relates to the technical field of electrochemical immunosensors, in particular to a construction method of a nanoprobe C60 based electrochemical immunosensor for carcino-embryonic antigens.The method includes: modifying gold nanoclusters on the surface of a glassy carbon electrode by means of cyclic voltammetry to obtain an antibody capture substrate; sequentially immobilizing first antibodies and antigens to the surface of the electrode; immobilizing AuNPs@L-cys-C60 marked second antibodies by specific binding of the antigens and the antibodies.The gold nanoclusters have excellent biocompatibility and electrical conductivity and large surface areas and are capable of promoting electron transferring between proteins and the electrode.The AuNPs@L-cys-C60 serves as an oxidation-reduction nanoprobe, the immunosensor is used for detection of the carcino-embryonic antigens on the basis of excellent specificity between the antigens and the antibodies, and detection of the carcino-embryonic antigens is realized according to different electrochemical signals of the carcino-embryonic antigens with different concentrations.

Description

technical field [0001] The invention relates to the technical field of electrochemical immunosensing, in particular to a nanoprobe-based C 60 Construction method of electrochemical immunosensor for carcinoembryonic antigen (CEA). Specifically involving a gold nanocluster as an antibody capture substrate, AuNPsL-cys-C 60 Construction method of electrochemical immunosensor with labeled secondary antibody as redox probe. Background technique [0002] CEA belongs to one of the embryonic antigens and is a normal component because it is produced by embryonic tissue during the embryonic development stage. With the growth of the embryo, the content of embryonic antigen in the later stage of the embryo decreases, and it will gradually disappear or only exist in a very small amount after birth. When cells become cancerous, such antigens can be re-synthesized and expressed on the surface of tumor cells, and can also be secreted into the blood, and their content is often positively c...

Claims

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Application Information

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IPC IPC(8): G01N27/48G01N27/327G01N33/574G01N33/543
CPCG01N27/48G01N27/3278G01N33/5438G01N33/57473
Inventor 郑香丽周长利夏方诠田栋花小霞乔雪莹
Owner UNIV OF JINAN
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