Preparation method and application of magnetic nanoparticles for capturing exosomes in blood

A technology of magnetic nanoparticles and micro-nanoparticles, which is applied in the field of tumor detection, can solve the problems of low processing capacity, long processing time, and low repetition rate, and achieve the effects of short processing time, small sample size, and easy operation

Active Publication Date: 2019-03-01
NINGBO MEIJING MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have many disadvantages based on their respective enrichment principles: the required sample size is large; the processing time is long; the repetition rate is low; the amount of inefficient processing is small, etc.

Method used

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  • Preparation method and application of magnetic nanoparticles for capturing exosomes in blood
  • Preparation method and application of magnetic nanoparticles for capturing exosomes in blood
  • Preparation method and application of magnetic nanoparticles for capturing exosomes in blood

Examples

Experimental program
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Effect test

Embodiment 1

[0020] A method for preparing magnetic nanoparticles for capturing exosomes in blood, comprising the following steps:

[0021] (1) 1.8mol ferric chloride hexahydrate (FeCl 3 ·6H 2 O) and 1 mol of ferrous chloride tetrahydrate (FeCl 2 4H 2 O) After mixing, dissolve in 100 mL of distilled water, stir and mix well, heat to 70°C, then slowly add NH with a concentration of 0.4mol / L 3 ·H 2 O solution, when the pH of the solution rises to 6-7, the iron salt hydrolyzes to produce a large amount of Fe 3 o 4 Crystal particles; continue to add NH dropwise 3 ·H 2 0 until the pH rises to 9-10, the mixed solution gradually turns black, and the hydrolysis tends to be complete; after the hydrolysis is basically completed, slowly add 2 mL of surfactant-cetyltrimethylammonium bromide (Cetyltrimethylammonium Bromide, CTAB), after stirring for 30 min, the product was collected by centrifugation; the product was repeatedly washed with distilled water until PH = 7.0, the supernatant was rem...

Embodiment 2

[0026] With above-mentioned embodiment 1, its difference is:

[0027] In step (1): mix ferric chloride hexahydrate and ferrous chloride tetrahydrate at a molar ratio of 1.6:1, dissolve in distilled water to obtain a mixed solution, stir and mix well, heat to 55°C, and then add dropwise at a concentration of 0.3mol / L of NH 3 ·H 2 O solution, when the hydrolysis of the mixed solution is completed, slowly add 1.5 mL of surfactant to the mixed solution, centrifuge to collect the product after stirring, wash the product repeatedly until pH = 7.0, remove the supernatant, take the lower layer and dry it in vacuum at 50 °C More than 16h, get nano-Fe 3 o 4 particle;

[0028] In step (2): take the nano-Fe obtained in step (1) 3 o 4 The particles were dispersed in a solution mixed with ethanol and distilled water at a ratio of 3:1 by volume. After ultrasonic treatment, a solution of tetraethyl orthosilicate with a concentration of 0.15 mol / L was added, and 6 mL was gradually added...

Embodiment 3

[0030] With above-mentioned embodiment 1, its difference is:

[0031] In step (1): mix ferric chloride hexahydrate and ferrous chloride tetrahydrate at a molar ratio of 2.0:1, dissolve in distilled water to obtain a mixed solution, stir and mix well, heat to 85°C, and then add dropwise at a concentration of 0.5mol / L NH 3 ·H 2 O solution, when the hydrolysis of the mixed solution is completed, slowly add 2.5 mL of surfactant to the mixed solution, centrifuge to collect the product after stirring, wash the product repeatedly until pH = 7.0, remove the supernatant, take the lower layer and dry it in vacuum at 70 °C More than 16h, get nano-Fe 3 o 4 particle;

[0032] In step (2): take the nano-Fe obtained in step (1) 3 o 4 The particles were dispersed in a solution mixed with ethanol and distilled water at a ratio of 5:1 by volume. After ultrasonic treatment, a solution of tetraethyl orthosilicate with a concentration of 0.25 mol / L was added, and 7.5 mL was gradually added ...

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Abstract

The invention discloses a preparation method and application of magnetic nanoparticles for capturing exosomes in blood. The preparation method is characterized by comprising the following steps: (1) after mixing diferric chloride hexahydrate and ferrous chloride tetrahydrate, dissolving a mixture into distilled water; after stirring and uniformly mixing, heating to 70 DEG C; then slowly dropwise adding an ammonia water solution; after hydrolysis is basically finished, slowly adding cetyl trimethyl ammonium bromide into the solution; stirring, centrifuging and collecting a product and washing;taking lower-layer sediment and drying in vacuum to obtain nano Fe3O4 particles; (2) after dispersing the nano Fe3O4 particles into an ethanol water solution and carrying out ultrasonic treatment, adding a tetraethyl orthosilicate solution; gradually adding into an ammonia water bath and reacting for 5 h; centrifuging and collecting a product; washing and drying to obtain the irregular magnetic micro-nanoparticles which are covered with SiO2 and have the surface distributed pore diameter of 40 to 200 nm. The preparation method disclosed by the invention has the advantages that the exosomes arelosslessly combined to the greatest extent, the amount of used samples is small, the treatment time is short and the operation is simple and convenient.

Description

technical field [0001] The invention belongs to the field of tumor detection, and in particular relates to a preparation method and application of magnetic nanoparticles for capturing exosomes in blood. Background technique [0002] In recent years, due to changes in the environment and lifestyle, the incidence of malignant tumors (also known as cancer) in my country has increased significantly. At the same time, the number of deaths from malignant tumors in my country is also increasing, which is largely due to the fact that most patients have reached the middle and late stages of cancer when they are clearly diagnosed, and have lost the best time for treatment. The survival rate of advanced cancer patients after treatment is extremely low, and the survival time is short, while the cure rate of early cancer patients is high, about 80%-90%. Therefore, early diagnosis is particularly important for cancer diagnosis and treatment. Currently, the diagnostic methods for maligna...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/10
Inventor 张晓晶沈挺宋毅
Owner NINGBO MEIJING MEDICAL TECH
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