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Bifunctional biomimetic enzyme/graphene oxide composite material and its preparation method and application

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 and high reactivity, and achieve the effect of good reproducibility

Active Publication Date: 2021-09-21
SUZHOU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 biomimetic enzyme/graphene oxide composite material and its preparation method and application
  • Bifunctional biomimetic enzyme/graphene oxide composite material and its preparation method and application
  • Bifunctional biomimetic enzyme/graphene oxide composite material and its preparation method and application

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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, keep stirring evenly, and then heat and reflux for 3 hours until the mixture turns dark blue, 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 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 catalysis of...

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 biomimetic enzyme / graphene oxide composite material, comprising: S1, FeCl 3 ·6H 2 O and PEI are soluble in K 3 Fe(CN) 6 , stirred, heated and refluxed for 2 to 5 hours, the resulting mixture was centrifuged and washed to obtain PEI@PB nanocubes; S2, MnSO 4 Dissolve DNA and water, heat to 50-80°C, then add K 3 PO 4 and water, stirred at 50-80°C for 1-2 hours, centrifuged to obtain DNA@Mn 3 (PO 4 ) 2 ; S3. Place the substrate coated with GO in the PEI@PB solution, shake for 2-3 hours, and obtain PEI@PB / GO under the catalysis of EDC and NHS; S4. In the PEI@PB / GO is coated with PDMS, heated at 60-90°C, and the PDMS / PEI@PB / GO is peeled off from the substrate; then the DNA@Mn 3 (PO 4 ) 2 Attached to the other side of GO, the bifunctional biomimetic enzyme / graphene oxide composite material is obtained. The invention also discloses the bifunctional biomimetic enzyme / graphene oxide composite material and its application as a sensor. The bifunctional biomimetic enzyme / graphene oxide composite material prepared by the present invention can be used to construct ROS sensors, which can detect H released by living cells in real time. 2 o 2 and O 2 ·‑ .

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 between 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 stab...

Claims

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Application Information

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Patent Type & Authority Patents(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