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Preparation method and application of immunosensor constructed based on copper nanocluster-resonance energy transfer system

A technology of resonance energy transfer and immune sensor, which is applied in the field of nanoscience and immune analysis, can solve the problem that the signal of the sensing platform is difficult to control, and achieve the effect of high sensitivity and wide detection range

Active Publication Date: 2019-12-03
UNIV OF JINAN
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  • Application Information

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Problems solved by technology

However, its high electroluminescence efficiency also makes it difficult to control the signal of the sensing platform. Based on this, we propose to use a resonance energy transfer system to control its luminescence intensity so that it can meet the needs of trace analysis of the target, aiming at The mechanism exploration of the extinct ECL sensing system provides a new way of thinking

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1. A kind of preparation method based on the immunosensor of copper nanocluster-resonance energy transfer system construction, is characterized in that, comprises the following steps:

[0026] (1) The glassy carbon electrode was ultrasonically cleaned in ethanol and deionized water for 30 s, respectively, and then washed with 1.0 μm, 0.3 μm and 0.05 μm Al 2 o 3 Polish it with polishing powder to make it as smooth as a mirror, and dry it with nitrogen;

[0027] (2) 6 μL uniformly dispersed Cu NCs-Au NPs solution with a concentration of 1 mg / mL was drop-coated on the surface of the glassy carbon electrode, and dried naturally at room temperature;

[0028] (3) Insert the above dried glassy carbon electrode into the 5 μg / mL recombinant protein A solution, at 4 o Incubate at C for 30 min, wash with ultrapure water, and dry naturally at room temperature;

[0029] (4) Add 6 μL of bovine serum albumin solution with a mass fraction of 0.1% dropwise to block non-spe...

Embodiment 2

[0032] Embodiment 2. A kind of preparation method of the immunosensor based on copper nanocluster-resonance energy transfer system construction, is characterized in that, comprises the following steps:

[0033] (1) The glassy carbon electrode was ultrasonically cleaned in ethanol and deionized water for 30 s, respectively, and then washed with 1.0 μm, 0.3 μm and 0.05 μm Al 2 o 3 Polish it with polishing powder to make it as smooth as a mirror, and dry it with nitrogen;

[0034] (2) 6 μL uniformly dispersed Cu NCs-Au NPs solution with a concentration of 2 mg / mL was drop-coated on the surface of the glassy carbon electrode, and dried naturally at room temperature;

[0035] (3) Insert the above-mentioned dry glassy carbon electrode into 7.5 μg / mL recombinant protein A solution, at 4 o Incubate at C for 30 min, wash with ultrapure water, and dry naturally at room temperature;

[0036] (4) Add 6 μL of bovine serum albumin solution with a mass fraction of 0.1% dropwise to block n...

Embodiment 3

[0039] Embodiment 3. A kind of preparation method based on the immunosensor of copper nanocluster-resonance energy transfer system construction, is characterized in that, comprises the following steps:

[0040] (1) The glassy carbon electrode was ultrasonically cleaned in ethanol and deionized water for 30 s, respectively, and then washed with 1.0 μm, 0.3 μm and 0.05 μm Al2 o 3 Polish it with polishing powder to make it as smooth as a mirror, and dry it with nitrogen;

[0041] (2) 6 μL uniformly dispersed Cu NCs-Au NPs solution with a concentration of 3 mg / mL was drop-coated on the surface of the glassy carbon electrode, and dried naturally at room temperature;

[0042] (3) Insert the above dried glassy carbon electrode into the 10 μg / mL recombinant protein A solution, at 4 o Incubate at C for 30 min, wash with ultrapure water, and dry naturally at room temperature;

[0043] (4) Add 6 μL of bovine serum albumin solution with a mass fraction of 0.1% dropwise to block non-spec...

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Abstract

The invention discloses a preparation method and application of an immunosensor constructed based on copper nano-cluster-resonance energy transfer system, and belongs to the field of nano-material andresonance energy transfer FRET. According to the method, copper nanoclusters Cu NCs coated by bovine serum albumin (BSA) serve as an energy donor, and gold nanoparticles Au NPs are grown in situ on the surface of the Cu NCs by utilizing L-ascorbic acid to serve as an energy receptor, and a Cu NCs-Au NPs three-dimensional resonance structure is obtained. Energy is transmitted between the donor-receptor pair through a dipole-dipole non-radiation action mode; the transmission rate is inversely proportional to the six power of the distance; and ECL emission of the Cu NCs is remarkably quenched and the luminous intensity is controlled within a reasonable range by virtue of high-ratio overlapping of spectra and ultra-short pairing distance between the two. The invention provides a new ECL-FRETenergy transfer model for trace detection of biomarkers, and provides a brand-new thought for mechanism exploration of a quenching type ECL sensing system.

Description

technical field [0001] The invention belongs to the field of nanometer science and immune analysis. Background technique [0002] Copper nanoclusters are a new type of nanomaterials emerging from the development of nanoscience. Its good optical stability, large Stokes shift, high quantum yield and good biocompatibility make it suitable for biosensing. The application has laid a solid foundation. Copper nanoclusters can be synthesized by various micro-reducing agents such as nucleic acids, ligands, and proteins in an alkaline environment. Among them, copper nanoclusters templated by bovine serum albumin exhibit short-wavelength electroluminescence at low potentials. At the same time, the bovine serum albumin coated on its surface exposes a large number of functional group active sites, which provides great convenience for the modification of biomolecules. Unique advantages. However, its high electroluminescence efficiency also makes it difficult to control the signal of th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/30G01N33/53
CPCG01N27/3278G01N27/308G01N33/53
Inventor 魏琴贾越杨磊薛经纬马洪敏赵磊
Owner UNIV OF JINAN
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