Folic acid functionalized and streptavidin modified magnetic nanoparticles, and preparation method and application thereof

A magnetic nanoparticle and streptavidin technology, which is applied in the preparation, sampling, and measurement devices of test samples, can solve the problems of inability to retain cell and molecular biology characterization and analysis, and achieves low cost and biocompatibility. good effect

Pending Publication Date: 2019-11-26
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This technique can only perform simple and preliminary techniques on circulating tumor cells, but cannot preserve the complete morphology of cells for further molecular biological characterization and analysis

Method used

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  • Folic acid functionalized and streptavidin modified magnetic nanoparticles, and preparation method and application thereof
  • Folic acid functionalized and streptavidin modified magnetic nanoparticles, and preparation method and application thereof
  • Folic acid functionalized and streptavidin modified magnetic nanoparticles, and preparation method and application thereof

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Embodiment 1

[0066] This example is used to illustrate the synthesis of magnetic nanoparticles of the present invention.

[0067] 1) 1.35gFeCl 3 ·6H 2 After O was dissolved in 40ml of ethylene glycol to form an orange-red transparent solution, 3.6g of anhydrous sodium acetate and 1.0g of anhydrous sodium acrylate were added, and heated at 50°C with vigorous stirring to form a khaki suspension and then ultrasonicated for more than 30 minutes. The suspension after sufficient sonication was transferred and sealed in a tetrapolyethylene fluoride stainless steel reaction kettle, and the temperature was slowly raised from room temperature to a reaction temperature of 200°C at a heating rate of 3°C / min and continued heating for 8 hours.

[0068] 2) After the product is separated from the liquid under the action of an external magnetic field with a magnetic field strength of 1000 kOe, it is washed three times with 50 ml of ethanol and 100 ml of ultrapure water to remove residual solvents and inor...

Embodiment 2

[0072] This example is used to illustrate the folic acid functionalization and characterization of the magnetic nanoparticles of the present invention.

[0073] 1) The solid content of 5mg magnetic nanoparticles is transferred to 1ml MES buffer solution (pH=6.5) after sufficient sonication; 10mg EDC (stored at -20°C) and 10mg sulfo-NHS (stored at 4°C) are placed at room temperature Afterwards, it was dissolved in the above-mentioned dispersion system of magnetic nanoparticles, and reacted with vigorous shaking at room temperature for 15 minutes.

[0074] 2) The activated magnetic nanoparticles were washed three times with 1 ml PBS to remove excess EDC, and then redispersed in a streptavidin solution with a concentration of 1.0 mg / ml dissolved in 1 ml PBS. The new magnetic nanoparticle dispersion was shaken and reacted at room temperature for 4 hours at the previous rotational speed of 1000 rpm. The modified magnetic nanoparticles were washed three times with 1 ml PBS and stor...

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Abstract

The invention provides a folic acid functionalized and streptavidin modified magnetic nanoparticles, which are prepared by modifying hydrophilic carboxylated superparamagnetic nanoparticles with streptavidin and then combining with biotin labeled folic acid molecules coupled with polyethylene glycol. The invention further provides a preparation method and application of the nanoparticles. The nanoparticles provided by the invention have low cost and good biocompatibility, and provide possibility for application in clinical blood samples. In addition, different from a traditional circulating tumor cell separation technology targeting epithelial cell adhesion molecules, the folic acid functionalized magnetic nanoparticles can capture tumor cells with limited epithelial cell adhesion moleculeexpression and high malignant degree, a rapid, precise and low-cost detection technology for enriching, separating and detecting circulating tumors is provided, and an effective method is provided for prognosis judgment, curative effect monitoring, metastasis and relapse monitoring, early detection and the like of clinical tumor metastasis patients.

Description

technical field [0001] The invention belongs to the technical field of liquid biopsy based on functionalized nanomaterials, and in particular relates to a folic acid functionalized streptavidin-modified magnetic nanoparticle, a preparation method and application thereof. Background technique [0002] Although circulating tumor cell capture technology based on targeting epithelial cell adhesion molecules has achieved some success, there is still the possibility of circulating tumor cell loss caused by the process of epithelial-mesenchymal transition, so there is an urgent need to establish an epithelial cell adhesion molecule complement Marker library. Folate receptor is a kind of glycosylphosphatidylinositol receptor, which is widely overexpressed in ovarian cancer, cervical cancer, triple negative breast cancer, colon cancer, non-small cell lung cancer and other tumor cells, and its expression level is related to cancer progression The degree is positively correlated, so i...

Claims

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Application Information

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IPC IPC(8): G01N1/40G01N1/38G01N1/34
CPCG01N1/4077G01N1/4022G01N1/38G01N1/34G01N2001/4094
Inventor 杨延莲郑望舒李平刘长亮王琛
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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