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A kind of method for preparing nanozyme probe

A nanozyme and probe technology, applied in chemical instruments and methods, instruments, physical/chemical process catalysts, etc., can solve the problems of high cost, unstable heme, etc., achieve low cost, improve sensitivity, and fast sensing time Effect

Active Publication Date: 2021-09-03
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But fixed heme carries the risk of instability and high cost

Method used

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  • A kind of method for preparing nanozyme probe
  • A kind of method for preparing nanozyme probe
  • A kind of method for preparing nanozyme probe

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Experimental program
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Effect test

Embodiment 1

[0050] Embodiment 1 nanozyme probe

[0051] The nanozyme probes of BNP-Hemin-AuNPs include Hemin-AuNPs nanozymes and BNP protein, where the transmission electron microscope images of Hemin-AuNPs nanozymes are shown in figure 1 , uniform size, smooth surface.

[0052] To evaluate the catalytic performance of Hemin-AuNPs nanozyme, UV-vis assay of ABTS oxidation reaction was performed. ABTS (10mM) was H 2 o 2 (0.5mM) oxidation to produce green cationic radical ABTS*. Therefore, the colorimetric signal can represent the catalytic activity of these enzymes. A blank control was set, the other control groups were Hemin-G4 group and AuNPs group, and the experimental groups were Hemin-AuNPs group and BNP-Hemin-AuNPs group.

[0053] The result is as figure 2 As shown in -A, the Hemin-AuNPs group has better catalytic activity than the Hemin-G4 group and the AuNPs group. Compared with the Hemin-AuNPs group, the BNP-Hemin-AuNPs group showed a slower catalytic rate but produced the ...

Embodiment 2

[0057] The preparation of the nanozyme probe of embodiment 2 BNP-Hemin-AuNPs

[0058] All glassware used was pre-filled with aqua regia (HCl:HNO 3 3:1 in volume ratio), rinse with ultrapure water and dry before use. Will pre-prepared 14mg·mL -1 PADS solution (14mg·mL -1 Concentration of pamidronate disodium salt aqueous solution) was quickly added to boiling 20mL HAuCl 4 In aqueous solution (1%), PADS: HAuCl 4 The mass ratio of the two is 5.0:1.0, and the observed solution color changes from light yellow to colorless, and then dark red. The solution was then cooled to reflux for 10 min and cooled to room temperature to obtain amino-surface-modified AuNPs (N-AuNPs). Amidation chemical synthesis method was used to couple hemin (Hemin) on N-AuNPs. That is, 300 μL of 1 mM Hemin solution (pH=11), 100 μL of Tween solution (1%), 15 mg of EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and 0.5 mg SH-PEG (mercapto-polyethylene glycol) was added to 10 mL N-AuNPs...

Embodiment 3

[0062] Example 3 Detection method of brain natriuretic peptide colorimetric sensing based on nanozyme probe

[0063]First, 1 mL of 0.22M EDC solution, 0.22M NHS solution (N-hydroxysuccinimide) and 10 mg / mL MNPs solution were mixed for 30 min under sonication. After magnetic separation, the first supernatant was removed, and the precipitate was washed with ultrapure water, and this step was repeated three times. Resuspend MNPs into solution using ultrapure water. 200 μL of BNP antibody solution (0.4 mM) was added to 3 mL of MNPs solution and stirred continuously for 120 min. After magnetic separation, the second supernatant was removed, and the precipitate was washed with ultrapure water. After repeating this step three times, the MNPs functionalized with the BNP antibody were resuspended as a solution, that is, the Anti-BNP-MNPs solution. Take 10 mL of the solution to be tested, add 1 mL each of the Hemin-BNP-AuNPs nanozyme probe solution and Anti-BNP-MNPs solution in Exampl...

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Abstract

The invention discloses a method for preparing a nanozyme probe. The nanozyme probe prepared by the method is used as a signal output means, which improves the sensitivity of biosensing, and also greatly improves the speed of biosensing so that The signal of biosensing can be output smoothly; an electrochemical sensing detection method is constructed based on the nanozyme probe, which has an extremely low detection limit and can detect trace amounts of BNP as low as fg level, reflecting a high degree of specificity and sensitivity, suitable for clinical and scientific research; a colorimetric chemical sensing detection method was constructed based on nanozyme probes, which has extremely fast sensing time and convenient operation, and can detect BNP quickly and easily, suitable for home use.

Description

technical field [0001] The invention relates to a nanozyme probe and a detection method for brain natriuretic peptide colorimetric / electrochemical sensing based on it. Background technique [0002] Brain natriuretic peptide (BNP) secreted by brain and ventricular cardiomyocytes is a 32 amino acid peptide. BNP plays a key role in body fluid balance. Recent studies have shown that elevated BNP has important implications for heart failure, differential diagnosis of various respiratory diseases, and assessment of acute myocardial infarction. The quantitative measurement and real-time tracking of BNP are of great significance for the treatment of diseases and the recovery of patients. Heart failure occurs primarily because the heart cannot pump blood to every part of the body through defects in the heart muscle. Because heart failure presents with a variety of symptoms, physicians face diagnostic and prognostic challenges. [0003] So far, the clinical detection of BNP is mai...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48B01J31/02B01J35/00
CPCB01J31/0244B01J35/0033B01J35/006G01N27/3272G01N27/48
Inventor 陈红霞
Owner SHANGHAI UNIV
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