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A kind of porous ceria nanorod composite structure and the preparation method of the enzyme solution based on the structure and the application of enzyme-linked immunoassay

A ceria and composite structure technology, applied in the field of nanomaterials, can solve the problems of large temperature influence on the catalytic ability of simulated enzymes, difficult practical application, etc., and achieve the effects of enhanced sensitivity, easy storage and transportation, and good reaction stability.

Inactive Publication Date: 2016-07-27
XI AN JIAOTONG UNIV
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Problems solved by technology

The materials studied above all have the disadvantage that the catalytic ability of simulated enzymes is greatly affected by temperature, which brings great difficulties to practical applications.

Method used

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  • A kind of porous ceria nanorod composite structure and the preparation method of the enzyme solution based on the structure and the application of enzyme-linked immunoassay
  • A kind of porous ceria nanorod composite structure and the preparation method of the enzyme solution based on the structure and the application of enzyme-linked immunoassay
  • A kind of porous ceria nanorod composite structure and the preparation method of the enzyme solution based on the structure and the application of enzyme-linked immunoassay

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Experimental program
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preparation example Construction

[0037] Second, the present invention provides a method for preparing a ceria nanorod simulated enzyme solution, comprising:

[0038] First mix the ceria nanorods, antigen or antibody, and coating buffer evenly, and incubate at a constant temperature; then centrifuge to remove free antigen or antibody to obtain ceria nanorods coated with antigen or antibody; finally Resuspended to prepare ceria nanorods simulated enzyme solution.

[0039] Among them, the pellet can be resuspended with deionized water or coating buffer. The concentration of ceria nanorods in the resuspended ceria nanorods simulated enzyme solution is 0.1-10mg / mL, such as 0.2mg / mL, 0.3mg / mL, 0.5mg / mL, 1mg / mL, 5mg / mL Or 10 mg / mL, more preferably 1 mg / mL.

[0040] Preferably, the coating buffer in the step is selected from a buffer solution containing bovine serum albumin (BSA); wherein, the buffer solution can be Tris buffer (Tris), phosphate buffer (PBS) and hydroxyl One or more of ethylpiperazine ethylsulfuri...

Embodiment 1

[0062] Embodiment 1: Preparation and purification of ceria nanorods

[0063] The ceria nanorods in the present invention can be obtained by methods well known to those skilled in the art. In the present invention, we firstly prepare the precursor of cerium oxide nanorods, and then obtain pure ceria nanorods by hydrothermal method.

[0064] Preparation of ceria nanorods: 5 mL of cerium nitrate aqueous solution (0.8 M) and 75 mL of NaOH aqueous solution (6.4 M) were mixed at room temperature for 30 min, and then hydrothermally treated at 100° C. for 24 h. After repeated washing with water and ethanol three times and drying, redisperse in water (2mg / mL), and hydrothermally treat at 180°C for 12h. Centrifuge at 12000rpm for 10min, remove the supernatant, and dry overnight at 60°C for later use.

[0065] TEM morphology of ceria nanorods and the corresponding specific surface area and Ce 3+ Scale maps such as figure 1 with figure 2 shown. from figure 1 It can be seen that th...

Embodiment 2

[0073] Embodiment 2: Enzyme kinetic parameter analysis

[0074] The reaction kinetics were investigated by monitoring the change of the absorbance of the oxidation product of 3,3',5,5'-tetramethylbenzidine. Use the continuous monitoring mode to measure every 1s interval. The test conditions were as follows: the concentration of ceria nanorods was 0.2 μg / mL, the concentration of HRP as a control was 1 ng / mL, 1.0 mL of 0.2M acetic acid buffer solution with pH 4.0, and the reaction temperature was set at 25°C. With 3,3',5,5'-tetramethylbenzidine as substrate, H 2 0 2 The concentration is fixed at 100mM; when H 2 o 2 As the substrate, the concentration of TMB was fixed at 0.8mM. The apparent kinetic parameters refer to the Lineweaver-Burk equation: l / V=(Km / V max )(1 / [C])+1 / V max get. where V is the reaction velocity, V max is the maximum reaction velocity, [C] is the substrate concentration, K m is the Michaelis constant.

[0075] The kinetic parameters obtained are sh...

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Abstract

The invention discloses a porous ceria nanorod composite structure and a preparation method of an enzyme solution based on the structure, as well as enzyme-linked immunosorbent assay (Elisa) application, and belongs to the technical field of nanomaterials. The porous ceria nanorod composite structure comprises a ceria nanorod and an antigen or an antibody coated on the outer surface of the ceria nanorod, wherein the simulated enzyme solution prepared by the ceria nanorod composite structure has a wide detection response range and high sensitivity against the antibody or the antigen, the detection limit can achieve the level of 10pg / mL, the preparation method is simple and easy to operate, and the Elisa detection method is easy to operate and good in environmental adaptability, and is not affected by environmental temperature.

Description

technical field [0001] The invention belongs to the technical field of nanomaterials, relates to porous ceria nanorods and applications thereof, in particular to a composite structure of porous ceria nanorods, a method for preparing an enzyme solution based on the structure, and an enzyme-linked immunoassay application. Background technique [0002] Enzyme-linked immunoassay (Elisa) is a widely recognized and powerful method for detecting proteins. The method has high sensitivity and standard operating procedures, and usually uses horseradish peroxidase (HRP)-labeled immunoreagents to generate signals to detect target molecules. Although it has the advantages of various methods and high sensitivity, it also has some disadvantages. For example, complex and expensive extraction processes are required, the temperature and pH stability of protease are poor, and the reaction process is highly dependent on temperature. In the past ten years, researchers have hoped for an Elisa m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/531B01J23/10
CPCB01J23/10B01J35/10G01N33/54346
Inventor 瞿永泉马媛媛田志敏
Owner XI AN JIAOTONG UNIV
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