Nano enzyme with nanometer cage core of hollow gold-platinum alloy and shell of porous silicon dioxide, preparation method and application thereof

A gold-platinum alloy and silicon dioxide technology, which is applied in the field of nano-enzymes and gold-platinum alloy core-silica shell nano-enzymes, can solve the problems of low catalytic efficiency of nano-enzymes, and achieves overcoming easy deactivation, extending the working environment and responding wide range of effects

Active Publication Date: 2019-02-19
ZAOZHUANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In view of the current lack of nano-enzyme-labeled antigen complexes and the low catalytic efficiency of existing nano-enzymes, the present invention provides a hollow gold-platinum alloy nano-cage core-porous silica shell nano-enzyme, which can be used for the compounding of labeled antigens substances, wide response range and high sensitivity in enzyme-linked immunoassay detection

Method used

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  • Nano enzyme with nanometer cage core of hollow gold-platinum alloy and shell of porous silicon dioxide, preparation method and application thereof
  • Nano enzyme with nanometer cage core of hollow gold-platinum alloy and shell of porous silicon dioxide, preparation method and application thereof
  • Nano enzyme with nanometer cage core of hollow gold-platinum alloy and shell of porous silicon dioxide, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Preparation of hollow gold-platinum alloy nanocage core-porous silica shell nanozyme

[0034] (1) Preparation of gold seeds

[0035] Take 7.5 mL of 0.1 M hexadecyltrimethylammonium bromide aqueous solution, add 100.4 μL of 24.7 mM tetrachloroauric acid aqueous solution to it, mix well and dilute the volume to 9.4 mL, under the condition of magnetic stirring Add 0.6 mL of sodium borohydride aqueous solution with a concentration of 0.01 M (temporarily prepared and placed in ice water before use), stop stirring for 3 min, and let it stand for 2 h to obtain a gold seed solution containing gold seeds. The gold seed solution The gold concentration was 0.25 mM.

[0036] (2) Preparation of gold nanorods

[0037] Take 100 mL of 0.1 M cetyltrimethylammonium bromide aqueous solution, add 2.05 mL of 24.7 mM tetrachloroauric acid aqueous solution, 1 mL of 10 mM silver nitrate aqueous solution and 2 mL of 0.5 M sulfuric acid aqueous solution, after mixing evenly, then ad...

Embodiment 2

[0044] Example 2 Preparation of Nanozyme-labeled Rubella Antigen Complex

[0045] (1) Preparation of hollow gold-platinum alloy nanocage core-porous silica shell nanoenzyme-labeled rubella antigen complex

[0046] Take 50 μL of hollow gold-platinum alloy nanocage core-porous silica shell solution (5nM), and disperse it in 450 μL of phosphate buffer solution (0.1M, pH 7) containing 50 μL of 10 mg / mL rubella virus antigen. After mixing evenly, put them in a 37°C incubator, and after incubation for 96 h, centrifuge at 12000 rpm for 5 min to remove free rubella virus antigen molecules. Pour off the supernatant and disperse the precipitate in 100μL phosphate buffer (0.1M, pH 7) for later use. Hollow Au-Pt alloy nanocage prepared by zeta potential characterization, hollow Au-Pt alloy nanocage core-porous silica shell nanozyme and hollow Au-Pt alloy nanocage core-porous silica shell nanoenzyme labeling Rubella antigen Compounds such as image 3 shown. The results showed that the ...

Embodiment 3

[0049] Example 3 Application of nano-enzyme-labeled antigen complex in enzyme-linked immunoassay

[0050] First, 300 μL of mouse anti-artificial immunoglobulin antibody at a concentration of 1 μg / mL was added to each well of a high-affinity 96-well enzyme-labeled microwell plate, and the coating buffer was 0.1 M phosphate buffer (pH=7 ) at 4 °C overnight, then the above-mentioned microplate was washed 5 times with phosphate-Tween buffer, and 300 μL of blocking solution (1.0 wt% bovine albumin) was added to each well to block at 37 °C for 2 After h, the cells were washed 5 times with phosphate-Tween buffer. Then, add 100 μL of a series of serum samples and incubate at 37 °C for 1 h, wash 5 times with phosphate-Tween buffer, then add 50 μL of nano-enzyme-labeled antigen complex and incubate for 0.5 h, so as to capture on the microwell plate The virus immunoglobulin antibody of the sample to be tested forms a primary antibody-secondary antibody-enzyme-labeled antigen complex, an...

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Abstract

The invention provides a nano enzyme with a nanometer cage core of a hollow gold-platinum alloy and a shell of a porous silicon dioxide, a preparation method and an application thereof, and belongs tothe field of nanometer materials. The preparation method comprises the following steps: etching a gold core platinum shell nanostructure through potassium hexachloroplatinate to obtain a nanocage ofhollow gold-platinum alloy; coating silicon dioxide on the surface of the nanocage through tetraethyl silicate; and obtaining the nano enzyme taking the nanometer cage, in a round rod shape, of the hollow gold-platinum alloy as a core, and a silicon dioxide shell as the periphery of the core. The length of the core is 80 to 100 nm, and the diameter is 20 to 25 nm. The shell is provided with a channel of 3 to 5 nm. The nano enzyme is negatively charged. The antigen can be bound by an attractive effect of positive and negative electric charges to form a nano enzyme label antigen complex, which is used for performing enzyme linked immunoassay. According to the nano enzyme with the nanometer cage core of the hollow gold-platinum alloy and the shell of the porous silicon dioxide, the preparation method and the application thereof, the cost of a reagent of the natural enzyme label antigen complex is reduced, the working environment of enzyme linked immunodetection is extended, the defects that the natural enzyme is easy to deactivate and denature, etc., are overcame, and the nano enzyme can be widely applied to various enzyme linked immunodetection.

Description

technical field [0001] The invention belongs to the field of nanomaterials, in particular to a nanozyme, in particular to a gold-platinum alloy core silica-shell nanozyme. Background technique [0002] Enzyme-Linked ImmunoSorbent Assay (ELISA) is the most widely used type of serum immunoassay, based on the immobilization of antigen or antibody and enzyme labeling of antigen or antibody. The antigen or antibody combined on the surface of the solid phase carrier still maintains its immunological activity, and the enzyme-labeled antigen or antibody retains both its immunological activity and enzyme activity. After adding the substrate of the enzyme reaction, the substrate is catalyzed by the enzyme to become a colored product, and the amount of the product is directly related to the amount of the test substance in the sample, so qualitative or quantitative analysis can be performed according to the color change of the reaction system. The enzyme-linked immunoassay method is co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/536G01N33/569
CPCG01N33/535G01N33/536G01N33/56983
Inventor 刘建波龙琳吴晓春
Owner ZAOZHUANG UNIV
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