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Preparation method of nanometer electrochemical aptasensor for detecting stress-induced phosphoprotein

An aptamer sensor and electrochemical technology, which is applied in the field of nano-electrochemical aptasensor preparation, can solve the problems that MB-DNA/Mn-MOF nanocomposites have not yet been retrieved, and achieve good specificity, high sensitivity, and easy operation. convenient effect

Active Publication Date: 2020-05-08
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no domestic and foreign literature and patent reports have been retrieved about MB-DNA / Mn-MOF nanocomposites and the construction of nano-electrochemical aptasensors based on the complexes for the quantitative detection of STIP1

Method used

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  • Preparation method of nanometer electrochemical aptasensor for detecting stress-induced phosphoprotein
  • Preparation method of nanometer electrochemical aptasensor for detecting stress-induced phosphoprotein
  • Preparation method of nanometer electrochemical aptasensor for detecting stress-induced phosphoprotein

Examples

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Effect test

Embodiment 1

[0016] The preparation method of the nano-electrochemical aptasensor based on the MB-DNA / Mn-MOF / GCE sensing interface involved in this example and the principle schematic diagram of its quantitative detection of STIP1, as shown in figure 1 As shown, the specific steps are as follows:

[0017] Preparation of manganese-doped nickel-based metal framework compound (Mn-MOF): 0.465gNi(NO 3 ) 2 ·6H 2 O and 0.01gMn(NO 3 ) 2 4H 2 O was dissolved in 25mL of dimethylformamide (DMF), and stirred for 30min to form solution A; 0.133g of 1,4-benzenedicarboxylic acid was dissolved in 25mL of DMF, and stirred for 30min to form solution B; under magnetic stirring, the solution B was slowly added to solution A, and then dilute hydrochloric acid was added dropwise to obtain a green solution, which was transferred to a high-pressure reactor and stirred at 180°C for 24 hours; the reaction product was centrifuged to obtain a precipitate, which was washed 3 times with DMF and ethanol , the prod...

Embodiment 2

[0021] The preparation method of the nano-electrochemical aptasensor based on the MB-DNA / Mn-MOF / GCE sensing interface involved in this example and the schematic diagram of the principle of quantitative detection of STIP1, and the preparation steps of Mn-MOF are the same as those in Example 1, other details The preparation steps are as follows:

[0022] Construction of sensing interface of methylene blue end-labeled single-stranded nucleic acid aptamer / manganese-doped nickel-based metal framework compound / glassy carbon electrode (MB-DNA / Mn-MOF / GCE): drop coating on clean glassy carbon electrode (GCE) surface 5 μL of Nafion solution (5% w / w), after incubation for 5 minutes, drop-coat Mn-MOF aqueous dispersion, where the concentration of Mn-MOF is 0.4 mg / mL, and the amount of drop-coating is 20 μL; MB-DNA (2 μM) at 37 ° C Incubate for 2 h, measure 15 μL and drop-coat on the surface of Mn-MOF / GCE, after incubation for 12 h, wash with N 2 Blow dry, then rinse with twice distilled ...

Embodiment 3

[0025] The preparation method of the nano-electrochemical aptasensor based on the MB-DNA / Mn-MOF / GCE sensing interface involved in this example and the schematic diagram of the principle of quantitative detection of STIP1, and the preparation steps of Mn-MOF are the same as those in Example 1, other details The preparation steps are as follows:

[0026] Construction of sensing interface of methylene blue end-labeled single-stranded nucleic acid aptamer / manganese-doped nickel-based metal framework compound / glassy carbon electrode (MB-DNA / Mn-MOF / GCE): drop coating on clean glassy carbon electrode (GCE) surface 5 μL of Nafion solution (5% w / w), after incubation for 5 minutes, drop-coat Mn-MOF aqueous dispersion, in which the concentration of Mn-MOF is 0.8 mg / mL, and the amount of drop-coating is 30 μL; MB-DNA (2 μM) at 37 ° C Incubate for 2 h, measure 20 μL and drop-coat on the surface of Mn-MOF / GCE, after incubation for 12 h, wash with N 2 Blow dry, then rinse with twice distill...

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Abstract

The invention discloses a method for preparing a nano-electrochemical aptamer sensor for detecting stress-induced phosphoproteins. A manganese-doped nickel-based metal framework compound Mn-MOF is prepared by a solvothermal method and drip-coated on the surface of a glassy carbon electrode GCE. , and then drop-coated methylene blue end-labeled single-stranded nucleic acid aptamer MB-DNA to construct the MB-DNA / Mn-MOF / GCE sensing interface. Adding the stress-induced phosphoprotein STIP1, the MB-DNA aptamer specifically binds to STIP1 to form a MB-DNA / STIP1 complex, which detaches MB-DNA from the surface of GCE and causes the peak intensity of MB oxidation current to weaken. The linear relationship between MB current peak intensity and STIP1 concentration was fitted to construct a nano-electrochemical aptasensor for detecting STIP1. The method of the invention is convenient to operate, has high sensitivity and good specificity, and can be used as a new method for quantitative detection of STIP1.

Description

technical field [0001] The invention belongs to the technical field of preparation of functional nanocomposite materials and electrochemical aptamer sensors, and in particular relates to a preparation method of a nanometer electrochemical aptamer sensor for detecting stress-induced phosphoproteins, and the prepared sensor can be used in biomedical samples for application. Sensitive and selective detection of phosphoproteins. Background technique [0002] Stress-induced phosphoprotein (STIP1) is a protein secreted by ovarian cancer tissue into the peripheral blood of patients. Detection of STIP1 can help improve the early diagnosis of ovarian cancer. In recent years, studies on STIP1 as a potential tumor marker for detecting human ovarian cancer have been reported successively. Huang et al prepared a gold nanoparticle-based hybrid nanocomposite regulated fluorescence on-off for the detection of human STIP1 (Yue Huang, Hao Li, Lei Wang, Xiaoxia Mao, Genxi Li, Highly sensitive...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/308G01N27/3277G01N27/3278
Inventor 桂日军孙泽君金辉
Owner QINGDAO UNIV
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