Preparation method of detection probe for alpha-synuclein accumulation as well as product and application of detection probe

A synuclein and detection probe technology, applied in the field of biological detection, to achieve the effects of small particle size, high biological safety, and good catalytic activity

Inactive Publication Date: 2018-12-14
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many studies have shown the superiority of these detection methods, there is still a lack of a method that can slow down the further aggregation of alpha-synuclein on the basis of effective detection

Method used

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  • Preparation method of detection probe for alpha-synuclein accumulation as well as product and application of detection probe
  • Preparation method of detection probe for alpha-synuclein accumulation as well as product and application of detection probe
  • Preparation method of detection probe for alpha-synuclein accumulation as well as product and application of detection probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Magnetic Fe3O4 nanoparticles (Fe 3 o 4 NPs) particle generation and surface amination modification.

[0034] Synthesis of Particle Size Fe by Chemical Co-precipitation 3 o 4 NPs: Weigh 0.6 g of FeCl 3 and 0.38 g FeSO 4, dissolved in 50 mL deionized water. 20 mL of 1.5 M NaOH was quickly added to the iron-containing solution, and stirred at room temperature for 10 min. The black precipitate was washed with ethanol and dried under vacuum overnight. Weigh an appropriate amount of Fe 3 o 4 NPs were characterized by XRD, with figure 1 It can be seen that the 2θ values ​​are 30.3︒, 35.7︒, 43.5︒, 53.9︒, 57.5︒, and 63.2︒, and there are 6 obvious diffraction peaks, whose positions are similar to those of Fe 3 o 4 The NPs can be well matched, corresponding to the (220), (311), (400), (422), (511) and (440) crystal planes of the cubic crystal phase, respectively.

[0035] Fe 3 o 4 NPs@SiO 2 Synthesis: 2.0 mg Fe 3 o 4 NPs were dispersed in 10 mL of cyclohe...

Embodiment 2

[0041] 1. Magnetic Fe3O4 nanoparticles (Fe 3 o 4 NPs) particle generation and surface amination modification.

[0042] Synthesis of Particle Size Fe by Chemical Co-precipitation 3 o 4 NPs: 0.6 g of FeCl 3 and 0.38g FeSO 4 , dissolved in 50 mL deionized water. 20 mL of 1.5 M NaOH was quickly added to the iron-containing solution, and stirred at room temperature for 10 min. The black precipitate was washed with ethanol and dried under vacuum overnight.

[0043] Fe 3 o 4 NPs@SiO 2 Synthesis: 2.0 mg Fe 3 o 4 NPs were dispersed in 10 mL of cyclohexane at room temperature, followed by 1.8 g Triton-X 100, 1.6 mL n-hexanol and 0.34 mL H 2 O was added to the above solution with stirring to form an inverse microemulsion. After 15 min, add 40 μL TEOS to the microemulsion, and after 30 min, add 0.1 mL of 28% ammonia water to the above mixture. After 24 h, ethanol precipitates Fe 3 o 4 NPs@SiO 2 Nanoparticles. Fe 3 o 4 NPs@SiO 2 The nanoparticles were separated by...

Embodiment 3

[0048] 1. Magnetic Fe3O4 nanoparticles (Fe 3 o 4 NPs) particle generation and surface amination modification.

[0049] Synthesis of 200 nm Fe by Hydrothermal Method 3 o 4 NPs: 1.35g FeCl 3 Dissolve in 40 mL ethylene glycol and stir evenly to form a bright yellow solution. 3.6 g Na Ac and 1.0 g PEG were added to the solution and stirred at room temperature for 30 min. The mixed solution was transferred into a reaction kettle and reacted at 200°C for 18 hr. The resulting black precipitate was washed with ethanol and dried under vacuum overnight.

[0050] Fe 3 o 4 NPs@SiO 2 Synthesis: 2.0 mg Fe 3 o 4 NPs were dispersed in 10 mL of cyclohexane at room temperature, followed by 1.8 g Triton-X 100, 1.6 mL n-hexanol and 0.34 mL H 2 O was added to the above solution with stirring to form an inverse microemulsion. After 15 min, add 40 μL TEOS to the microemulsion, and after 30 min, add 0.1 mL of 28% ammonia water to the above mixture. After 24 h, ethanol precipitates F...

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Abstract

The invention relates to a preparation method of a detection probe for alpha-synuclein accumulation as well as a product and application of the detection probe. According to the preparation method, magnetic ferroferric oxide nanoparticles (Fe3O4 NPs) are used as carriers, and tetraethyl orthosilicate (TEOS) and 3-aminopropyltrimethoxysilane (APTES) are used for aminated modification of the surfaceof Fe3O4 NPs; further, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) are used as coupling agents for binding alpha-synuclein monoclonal antibodies; the Fe3O4 NPscarriers are coated with silica shell by utilizing the autocatalytic activity of Fe3O4 NPs, and then the alpha-synuclein monoclonal antibodies are bound to synthesize the detection probe which targets a Parkinson's disease, and is capable of fixed-point clearing intracellular ROS, and delaying the further accumulation of alpha-synuclein. The detection probe combines targeting, imaging and treatment into one, and is simple in synthesis method; the raw materials used are high in biosafety, relatively good in catalytic activity and good in physical stability.

Description

technical field [0001] The invention relates to a preparation method of a detection probe for α-synuclein accumulation, as well as its product and application. The invention belongs to the field of biological detection. Background technique [0002] Parkinson's disease is a common neurodegenerative disease in the elderly. Its main clinical features are resting tremor, slow movement, muscle rigidity, and abnormal posture and gait, which brings great pain to patients and their families. In patients with Parkinson's disease, a large number of degeneration and loss of dopamine neurons in the "substantia nigra" of the midbrain are accompanied by the formation of a large number of Lewy bodies in the neuron cytoplasm. Studies have shown that the accumulation of α-synuclein is closely related to the formation of Lewy bodies. Alpha-synuclein is a 140-amino acid, abundant brain protein that lacks cysteine ​​and tryptophan residues. It exists at the end of nerve cells and is mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577
CPCG01N33/54326G01N33/54346G01N33/577
Inventor 何丹农徐艳陈玮嘉张兆坤王萍金彩虹
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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