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Bifunctional bionic enzyme/graphene oxide composite as well as preparation method and application thereof

A composite material and biomimetic enzyme technology, applied in the field of bifunctional biomimetic enzyme/graphene oxide composite materials, can solve the problems of short biological half-life, low concentration, high reactivity, etc., and achieve good reproducible results

Active Publication Date: 2019-12-20
SUZHOU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its extremely short biological half-life, high reactivity, and low concentration, the in situ real-time detection of ROS is still a great challenge.

Method used

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  • Bifunctional bionic enzyme/graphene oxide composite as well as preparation method and application thereof
  • Bifunctional bionic enzyme/graphene oxide composite as well as preparation method and application thereof
  • Bifunctional bionic enzyme/graphene oxide composite as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Synthesis of PEI@PB / GO / DNA@Mn 3 (PO 4 ) 2 composite material

[0033] First, 10.0 mL of 5 mM FeCl 3 ·6H 2 O (pH=1.1) and 1.0 mL of 3% PEI were added to 10 mL of 5 mM K 3 Fe(CN) 6 (pH=1.1) solution, continue to stir evenly, and then heat and reflux for 3 hours until the mixture turns dark blue, and then wash with deionized water several times and centrifuge to obtain PEI@PB nanocubes. Second, mix 2.1mg DNA and 10.0mL 0.1M MnSO 4 The solution was added to 9mL secondary water and heated to 60°C, then 1.0mL 0.1M K 3 PO 4 Add the above mixed solution with 9mL of secondary water, then heat the mixed solution to 60°C and keep stirring for 1 hour, and then centrifuge to obtain DNA@Mn 3 (PO 4 ) 2 . Furthermore, GO was first prepared by the improved Hummers method, and then 0.5 mg / mL GO aqueous dispersion was deposited on the glass substrate by spin coating method, and then the substrate was placed in PEI@PB solution for 2 hours, and then 100 mM Under the ...

Embodiment 2

[0034] Embodiment 2: the preparation of ROS detection sensor

[0035] First, the glassy carbon electrode (GCE) was polished with 0.3 μm and 0.05 μm alumina powder, and then ultrasonically cleaned in deionized water, absolute ethanol, and deionized water for 3 min, respectively. After the electrode surface was dry, 20 μL prepared PEI@PB / GO / DNA@Mn 3 (PO 4 ) 2 The solution is dripped onto the electrode surface. Finally, the modified electrode was dried at room temperature for 10 h.

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Abstract

The invention discloses a preparation method of a bifunctional bionic enzyme / graphene oxide composite. The preparation method comprises the following steps: S1, dissolving FeCl3.6H2O and PEI into K3Fe(CN)6, stirring, heating and refluxing for 2-5 hours, and centrifuging and washing the obtained mixture, thus obtaining a PEI@PB nanocube; S2, dissolving MnSO4 and DNA into water, heating to 50-80 DEGC, then adding K3PO4 and water, stirring for 1-2 hours at the temperature of 50-80 DEG C, and centrifuging, thus obtaining DNA@Mn3(PO4)2; S3, placing a substrate coated with GO into PEI@PB solution,shaking for 2-3 hours, and under the catalytic action of EDC and NHS, obtaining PEI@PB / GO; and S4, coating PDMS on the PEI@PB / GO, heating at the temperature of 60-90 DEG C, and stripping PDMS / PEI@PB / GO from the substrate; and then attaching DNA@Mn3(PO4) to the other surface of the GO, so that the bifunctional bionic enzyme / graphene oxide composite is obtained. The invention also discloses the bifunctional bionic enzyme / graphene oxide composite and application thereof as a sensor. The bifunctional bionic enzyme / graphene oxide composite prepared by adopting the preparation method disclosed by the invention can be used for constructing an ROS sensor, and H2O2 and O2<.-> which are released by living cells can be detected in real time.

Description

technical field [0001] The invention relates to the technical field of biosensing, in particular to a bifunctional biomimetic enzyme / graphene oxide (GO) composite material, its preparation method and sensing application. Background technique [0002] Reactive oxygen species (ROS) are by-products of biological aerobic metabolism, which can mediate intracellular signal transmission and play an important role in regulating cell proliferation, differentiation, aging, and apoptosis. The imbalance of ROS production and elimination will cause oxidative stress, and excessive ROS will inhibit the normal life activities of cells. Therefore, ROS not only play an important role in many physiological processes, but also are related to many diseases including cancer, cardiovascular disease, neurological disease, diabetes, chronic inflammation and so on. Hydrogen peroxide (H 2 o 2 ) and superoxide anion (O 2 · - ) are the two main components of ROS. h 2 o 2 It is the most stable RO...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/308G01N27/3272
Inventor 胡芳馨郭春显杨鸿斌饶强海陈逸飞李长明胡光煊
Owner SUZHOU UNIV OF SCI & TECH