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A preparation method and application of magnetic nanoparticles for capturing exosomes in blood

A magnetic nanoparticle, micro-nanoparticle technology, applied in the field of tumor detection, can solve the problems of inefficient processing, long processing time, large sample volume, etc., and achieve the effects of short processing time, small sample volume, and easy operation.

Active Publication Date: 2022-02-22
NINGBO MEIJING MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A preparation method and application of magnetic nanoparticles for capturing exosomes in blood
  • A preparation method and application of magnetic nanoparticles for capturing exosomes in blood
  • A 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 mixture 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 removed, ...

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, which is characterized by the following steps: (1) mixing ferrous chloride hexahydrate and ferrous chloride tetrahydrate In distilled water, stir and mix well and heat to 70°C, then slowly add ammonia solution dropwise, after the hydrolysis is basically completed, slowly add cetyltrimethylamine bromide to the solution, stir and centrifuge to collect the product for washing, take the lower precipitate and dry it in vacuum to obtain Nano Fe 3 o 4 Particles; (2) take nano Fe 3 o 4 After the particles were dispersed in an aqueous ethanol solution and ultrasonically treated, after adding tetraethyl orthosilicate solution, they were gradually added to an aqueous ammonia water bath to react for 5 h, and the product was collected by centrifugation, washed and dried to obtain SiO coated particles. 2 In addition, irregular magnetic micro-nano particles with a pore size of 40-200 nm are distributed on the surface, which has the advantages of binding exosomes without damage to the greatest extent, and the sample volume used is small, the processing time is short, and the operation is simple.

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 Patents(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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