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Application of nano-enzyme in catalytic enhancement of electrochemical luminescence signals and single-cell surface film protein imaging

A luminescent signal and nanozyme technology, applied in the field of chemical analysis, can solve the problems of weak signal and low sensitivity of luminescent molecules

Inactive Publication Date: 2022-04-15
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide the application of nanozyme in catalyzing enhanced electrochemiluminescence signal and imaging of single cell surface membrane protein, Co 3 o 4 Nanozyme-enhanced electrochemiluminescence imaging for small biomolecules H 2 o 2 And high-sensitivity imaging analysis of single-cell surface membrane proteins, solving the problems of weak luminescent molecule signals and low sensitivity in existing electrochemiluminescence imaging

Method used

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  • Application of nano-enzyme in catalytic enhancement of electrochemical luminescence signals and single-cell surface film protein imaging
  • Application of nano-enzyme in catalytic enhancement of electrochemical luminescence signals and single-cell surface film protein imaging
  • Application of nano-enzyme in catalytic enhancement of electrochemical luminescence signals and single-cell surface film protein imaging

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Co 3 o 4 Synthesis and surface functionalization of nanoparticles

[0033] Weigh 1g Co(NO 3 ) 2 ·6H 2 O, dissolved in 40 mL of ultrapure water. Add 0.8 mL of 30% hydrogen peroxide solution. The pH of the solution was adjusted to 9.0 with aqueous ammonia. Subsequently, the solution was transferred to a reactor and reacted at 180 °C for 12 h. After the reaction, the product was centrifuged, washed 3 times, and dried to obtain Co 3 o 4 Nanoparticles. Will Co 3 o 4 Disperse in a certain amount of ultrapure water, let it stand for 3 hours, and centrifuge to obtain Co 3 o 4 Nanoparticles. Disperse it in a mixed solution of absolute ethanol and water, ultrasonically mix, and add 3-aminopropyltriethoxysilane. Stir overnight at 50 °C, centrifuge, wash, and dry to obtain the surface-aminated Co 3 o 4 .

[0034] to Co 3 o 4 Nanoparticles were characterized by TEM, as figure 2 As shown in A, Co 3 o 4 The particle size of the nanoparticles is about ...

Embodiment 2

[0035] Example 2 Co 3 o 4 Electrochemiluminescence performance analysis of nanozyme

[0036] Use an O-ring to fix a certain size conductive area on the ITO electrode as the working electrode. The surface aminated Co prepared in Example 1 3 o 4 Disperse on the indium tin oxide electrode, and dry at 80°C to spread the particles evenly and firmly on the surface of the electrode. will be paved with Co 3 o 4 Enzyme ITO electrode as working electrode, Ag / AgCl electrode as reference electrode, Pt wire as auxiliary electrode, L012-H 2 o 2 As a luminescent system, cyclic voltammetry scanning was performed at a scan rate of 0.1 V / s in the range of 0-1 V (vs. Ag / AgCl), and the ECL curve was detected and recorded on an ECL analysis instrument; compared with no Co 3 o 4 , the ECL signal of the ITO electrode ( figure 2 in B), it can be seen that Co 3 o 4 When present, L012-H 2 o 2 The ECL signal of the was significantly enhanced, indicating that the Co 3 o 4 It has excellen...

Embodiment 3

[0038] Example 3 Co 3 o 4 Application of Nanozyme in Electrochemical Sensing Analysis

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Abstract

The invention provides an application of a nano-enzyme in catalytic enhancement of an electrochemical luminescence signal and single-cell surface membrane protein imaging, the nano-enzyme is a Co3O4 nano-enzyme, an electrochemical luminescence system is luminol analogue L012-H2O2, and a membrane protein is a carcino-embryonic antigen (CEA). The Co3O4 catalysis enhanced ECL visualization is obtained for the first time, voltage is applied to a luminol analogue L012-H2O2 system, hydrogen peroxide is decomposed to generate reactive oxygen free radicals under the catalysis of nano-enzyme, L012 is enhanced to generate a light-emitting signal, and therefore a locally-enhanced visual ECL signal is generated on the nano-enzyme and is used for high-sensitivity imaging analysis of single-cell surface film protein. A CEA antibody is modified on Co3O4 through covalent binding to prepare the functionalized probe. By means of immune binding, the cells can specifically capture the probe. And hydrogen peroxide generates reactive oxygen free radicals under the catalysis of Co3O4.

Description

technical field [0001] The invention belongs to the field of chemical analysis, and in particular relates to the application of nanozymes in catalyzing and enhancing electrochemiluminescent signals and single-cell surface membrane protein imaging. Background technique [0002] Electrochemiluminescence is a light radiation phenomenon produced by chemical molecules undergoing an electrochemical reaction process. Because the electrochemiluminescence process relies on an external voltage to drive the luminescence process. Compared with fluorescence technology, electrochemiluminescence does not require an additional excitation light source, which effectively avoids the interference of light scattering on detection and improves detection sensitivity. Now it has become a powerful analytical technique widely used in biochemical analysis, pharmaceutical analysis, life science analysis, environmental analysis and other fields. In order to further improve the analytical throughput an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/416G01N27/30G01N33/574
Inventor 张晶晶晁洁
Owner NANJING UNIV OF POSTS & TELECOMM