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Construction method and application of platinum-shell gold-core nano-enzyme mediated magnetic relaxation sensing signal amplification system

A construction method and technology for sensing signals, applied in the fields of resources and environmental protection, can solve problems such as poor stability and lack of linear range magnetic signal probes

Pending Publication Date: 2021-09-10
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the target can only bind to a limited number of MNPs, lacks a sufficiently wide linear range and a sufficiently strong magnetic signal probe, and because the number and state of MNPs are easily interfered by many factors such as the sample matrix, resulting in non-specific aggregation, leading to the method less stable

Method used

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  • Construction method and application of platinum-shell gold-core nano-enzyme mediated magnetic relaxation sensing signal amplification system
  • Construction method and application of platinum-shell gold-core nano-enzyme mediated magnetic relaxation sensing signal amplification system
  • Construction method and application of platinum-shell gold-core nano-enzyme mediated magnetic relaxation sensing signal amplification system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. Preparation of gold core platinum shell nanoparticles (Pt@AuNPs)

[0068] 1) Preparation of gold nanoparticles (Gold Nanoparticle Seeds, AuNPs)

[0069] In order to avoid the impurity of the prepared nanoparticles, the utensils required for the experiment should be soaked with freshly prepared aqua regia (concentrated nitric acid with a volume ratio of 1:3 and concentrated hydrochloric acid: 1:3) before the experiment started. Soak for 3 h, rinsed with ultrapure water, and dried for use. 200 μL HAuCl 4 (48 μM) and 9.8 mL of ultrapure water were added to the reaction kettle, and kept boiling for 1 min under microwave heating at 100 °C. Then turn on the magnetic stirring, quickly add 1 mL of 1wt% sodium citrate at one time, keep at 100 °C for 4 min, and then cool to room temperature to obtain AuNPs, which are stored at 4 °C for later use.

[0070] 2) Preparation of Pt@AuNPsNPs

[0071] Pt@AuNPs were synthesized by platinum precipitation on the surface of Au...

Embodiment 2

[0076] Example 2. Performance evaluation of Pt@AuNPs

[0077] 1) Using peroxidase in H 2 o 2 In the presence of the catalytic substrate 3,3,5,5-tetramethylbenzidine (TMB), a blue reaction occurred to test the peroxidase activity of Pt@AuNPs. Add 100 μL of Pt@AuNPs suspension with different concentrations (0.225 pmol / L-0.46 nmol / L) to 100 μL multi-component TMB chromogenic solution to produce a blue reaction, and after 5 min, use H 2 SO 4 Quenched. Observe its color change and measure its UV absorbance (OD 650 ). Experimental results such as image 3 As shown, the absorbance at 650 nm increases with the increase of the concentration of Pt@AuNPs. It is proved that Pt@AuNPs have good catalase catalytic activity.

[0078]2) Comparison of Pt@AuNPs (0.89 pmol / L-0.92 nmol / L) and HRP (10 -2 ng / mL-10 3 ng / mL) with the storage time of its peroxidase activity changes, to determine the stability of Pt@AuNPs and HRP. Pt@AuNPs and HRP were stored at 4°C and room temperature (22...

Embodiment 3

[0079] Example 3. Pt@AuNPs-mediated magnetic relaxation immunosensor for the detection of procalcitonin

[0080] 1) MNPs-procalcitonin capture antibody (MNP S -Ab 1 ) Preparation of conjugates

[0081] Take 200 μL of MNP S (10 mg / mL) in a 1.5 mL centrifuge tube, put it on a magnetic separation rack, wash twice with MEST (10 mM MES, pH=6.0, 0.05 % Tween-20) and place it on a magnetic separation rack separate. Remove the supernatant and add 80 μL EDC (5 mg / mL) and 40 μL NHS (5 mg / mL) to the centrifuge tube. The mixture was then gently shaken on a vortex shaker at 37°C for 30 min. Then place the tube on a magnetic separation rack for separation, wash twice with MEST and resuspend, then add 50 μg of procalcitonin antibody to the tube, and adjust the volume to 500 μL with PBST. The mixture was shaken gently on a vortex shaker for 3 h at 37 °C. After magnetic separation and supernatant removal, add 1 mL of PBST containing 1% BSA to MNP s -Ab 1 In the conjugate, place at 37°...

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Abstract

The invention belongs to the technical field of biochemical analysis, and concretely relates to a construction method and application of a platinum-shell gold-core nano-enzyme mediated magnetic relaxation sensing signal amplification system. The biosensor is characterized in that gold nanoparticles are synthesized by a microwave synthesis method, Pt@AuNPs nano-enzyme is prepared by platinum deposition on the surfaces of the gold nanoparticles through an oxidation-reduction reaction between ascorbic acid and chloroplatinic acid by taking the gold nanoparticles as a core, the Pt@AuNPs nano-enzyme is used as a signal amplifier, a hydrogen peroxide mediated paramagnetic ion valence conversion sensing system is used as a magnetic relaxation signal reading system, and a Pt@AuNPs nano-enzyme mediated magnetic relaxation immunosensor and a magnetic relaxation DNA sensor are respectively constructed and are used for rapid and high-sensitivity detection of bacterial infection markers and food-borne pathogenic bacteria.

Description

technical field [0001] The invention relates to the technical field of resources and environmental protection, in particular to a construction method and application of a platinum-shell gold-core nanozyme-mediated magnetic relaxation sensing signal amplification system. Background technique [0002] In recent years, food safety problems have emerged one after another, causing serious health threats and economic losses to people in my country and the world. In 2003, the World Health Organization listed food safety as one of the top ten threats to global public health. Foodborne diseases caused by foodborne pathogens are one of the most prominent food safety problems, accounting for more than 45% of the total number of global food safety incidents. According to the statistical data of "China Health Statistical Yearbook", from 2015 to 2018, the number of people who fell ill due to food-borne pathogens in my country accounted for 29.3% of all food-borne illnesses. At the same t...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/68
CPCG01N33/543G01N33/6854
Inventor 陈翊平吴紫荆董永贞
Owner HUAZHONG AGRI UNIV
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