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Preparation and application of adjacent hybridization dual-mode immunosensor based on MXene nanosheet photo-thermal amplification

An immunosensor and nanosheet technology, which can be applied to anti-peptide structure protease inhibitor immunoglobulins, instruments, scientific instruments, etc., and can solve the problems of limited application of instrument analysis technology.

Active Publication Date: 2020-01-21
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, in the clinical diagnosis of tumor markers, a variety of methods based on proximity hybridization strategies have been developed, and the application of traditional instrumental analysis techniques is limited in some resource-constrained areas

Method used

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  • Preparation and application of adjacent hybridization dual-mode immunosensor based on MXene nanosheet photo-thermal amplification
  • Preparation and application of adjacent hybridization dual-mode immunosensor based on MXene nanosheet photo-thermal amplification
  • Preparation and application of adjacent hybridization dual-mode immunosensor based on MXene nanosheet photo-thermal amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Fabrication of a proximity hybridization dual-mode immunosensor based on photothermal amplification of MXene nanosheets (e.g. figure 1 shown):

[0027] (1) Pretreatment of the glassy carbon electrode: the glassy carbon electrode is first mechanically polished and polished on the suede covered with alumina powder, washed with secondary water to remove the residual powder on the surface, and then moved into an ultrasonic water bath for cleaning until it is cleaned, and finally Wash thoroughly with ethanol, dilute acid and water;

[0028] (2) Add 3 mg of TiO 2 Discoid nanoparticles (TiO 2 NDs) powder was dissolved in 1 mL ultrapure water to obtain 3 mg / mLTiO 2 For the aqueous dispersion of NDs, take 10 µL polyethyleneimine (PEI) and dilute it with ultrapure water to 1 mL to obtain a 1 wt.% PEI aqueous solution, and add 1 mL of the above TiO 2 Add 1 mL of 1 wt.% PEI aqueous solution to the NDs dispersion, stir continuously at room temperature for 6 h, centrifuge at 1...

Embodiment 2

[0034] TiO used in embodiment 1 2 Discoid nanoparticles (TiO 2 NDs) Preparation:

[0035] Mix 4.5 g terephthalic acid, 2.34 mL butyl(IV) titanate, 9 mL anhydrous methanol, and 81 mL N,N-dimethylformamide (DMF), then transfer the mixture to a 100 mL autoclave , heated at 150 °C for 48 h, filtered after cooling and washed with methanol several times to obtain a white suspension, subsequently, the above-mentioned white suspension product was calcined in air at 400 °C for 5 h, after natural cooling, the white powder obtained was collected as TiO 2 Discoid nanoparticles (TiO 2 NDs);

[0036] The DNA used in Example 1 1 -Ab 1 or DNA 2 -Ab 2 Preparation of:

[0037] Antibody (Ab 1 or Ab 2 ) activation: take 0.2 mg antibody (Ab 1 or Ab 2 ) solid was diluted to 0.2 mg / mL with 1 mL of 55 mM, pH 7.4 phosphate buffer solution, and 10 µL of 0.4 mM (N-maleimidomethyl)cyclohexane-1-carboxylate succinyl Dimethyl sulfoxide solution of imidate (SMCC), mixed with 100 µL of 0.2 mg...

Embodiment 3

[0041] An application of a proximity hybridization dual-mode immunosensor based on photothermal amplification of MXene nanosheets, the steps are as follows:

[0042] (1) The three-electrode system was used for measurement, and TiO 2 NDs-PEI / DNA 3 / DNA 1 -Ab 1 -HE4-DNA 2 -Ab 2 The / MXene@Thi modified electrode was used as the working electrode, Ag / AgCl was used as the reference electrode, and the platinum wire electrode was used as the auxiliary electrode. HE4 was detected on the electrochemical workstation, and the initial potential was set to 0 V, and the termination potential was set to -0.5 V;

[0043] (2) In a phosphate buffer solution of pH 7.4, square wave voltammetry (SWV) was used to measure 1×10 -6 ng / mL~10 ng / mL series of HE4 standard solutions with different concentrations were used to measure the electrochemical behavior on the modified electrode. By recording the electrochemical signal generated after 808 nm laser irradiation, the working curve was drawn, s...

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Abstract

The invention discloses preparation and application of an adjacent hybridization dual-mode immunosensor based on MXene nanosheet photo-thermal amplification. Polyethyleneimine (PEI) modified TiO2 disc-shaped nanoparticles are used as a substrate, and a large amount of capture substrate deoxyribonucleic acids are fixed on a sensing interface through electrodeposition of gold. Two deoxyribonucleic acid probes are both labeled with a human epididymis protein 4 (HE4) antibody of a target object, and after the deoxyribonucleic acid probes as a recognition part of HE4 are captured by DNA3, the HE4 induces adjacent hybridization between primary antibody-labeled DNA1 and secondary antibody-labeled DNA2 by immunorecognition. An MXene nanosheet is loaded with a large amount of thionine, so that theMXene nanosheet can be effectively intercalated into a double-stranded DNA groove formed by hybridization, can be used as a signal probe for electrochemical-temperature dual modes, and realizes amplification of electrochemical signals due to temperature rise generated by the photo-thermal effect. Under the irradiation of an 808 nm infrared laser device, high-sensitivity detection of the HE4 is realized.

Description

technical field [0001] The invention belongs to the technical field of novel functional materials and biosensing detection, and specifically relates to the preparation and application of a proximity hybridization dual-mode immune sensor based on photothermal amplification of MXene nanosheets. Background technique [0002] As a special analytical strategy, protein-binding deoxyribose nucleic acid (DNA)-mediated proximity hybridization has received great attention in the identification and detection of proteins. After a pair of DNA-labeled affinity probes go through the process of immune reaction and DNA hybridization, the target protein is recognized simultaneously, which greatly enhances the selectivity and sensitivity of the target protein, which is attributed to the protein and the DNA of the labeled probe. Double identification. Therefore, this method converts traditional protein immune recognition into simple DNA recognition, significantly simplifies the operation proce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/68G01N33/574G01N27/30C07K16/38
CPCG01N33/531G01N33/68G01N33/57449G01N27/30C07K16/38G01N2333/81
Inventor 戴宏皇怡甜任卉竹高利红张书培
Owner FUJIAN NORMAL UNIV
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