Magnetic bead for capturing circulating tumor cells and preparation method thereof

A tumor cell, magnetic bead technology, applied in the field of biomedical engineering, can solve the problems of the influence of cell biochemical characteristics, unfavorable cell detection, etc., to achieve good biocompatibility and stability, efficient separation and capture effect, high degree of dispersion Effect

Pending Publication Date: 2020-11-06
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current method uses magnetic beads to directly bind CTC, which will have a certain impact on the b

Method used

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  • Magnetic bead for capturing circulating tumor cells and preparation method thereof
  • Magnetic bead for capturing circulating tumor cells and preparation method thereof
  • Magnetic bead for capturing circulating tumor cells and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0043] Example 1 Preparation of Magnetic Beads

[0044] according to figure 1 and figure 2 As shown in the procedure, the magnetic beads of the present invention were prepared.

[0045] Silanized Fe 3 O 4 Magnetic beads, the preparation method is a conventional method, and its modification can be 3-aminopropyltriethoxysilane (APTMS), 3-mercaptopropyltrimethoxysilane (MPTMS).

[0046] Colloidal gold is prepared by redox method, and its preparation method can be as follows:

[0047] Dissolve tetrachloroalloy acid, deionized water and trisodium citrate in deionized water, and stir mechanically for 2 minutes to obtain a colloidal gold pre-solution. The sodium borohydride is dissolved in the trisodium citrate solution, the colloidal gold pre-solution is added, the glass rod is rapidly stirred for 5 minutes to obtain the colloidal gold, and the prepared colloidal gold particles have a diameter of 20nm-30nm.

[0048] The preparation method of magnetic beads includes the follow...

Example Embodiment

[0059] Example 2

[0060] The magnetic beads prepared in Example 1 for capture were dissolved in absolute ethanol at a concentration of 1 g / L, using Malvern The potentiometer measures the zeta potential of the colloid surface. The results show that the surface potential of the colloidal system is -63.6mV, which indicates that the solution has excellent stability at a temperature of 25.0℃. The results are as follows image 3 shown.

Example Embodiment

[0061] Example 3

[0062] The gold-coated surface in the culture medium was modified in the same way as the surface of the magnetic beads for capture prepared by the present invention to simulate and verify the effect of the magnetic beads for capture on cells. The specific methods include:

[0063] In a circular petri dish, the same proportion of MCH and MUA anhydrous ethanol solution as the magnetic bead modification in Example 1 was prepared, and the gold foil-sputtered coverslip was soaked overnight, and then in a pH 7.4 PBS solution according to the example The weight ratio of NHS / EDC in 1 was used for NHS / EDC coupling reaction and binding anti-EpCAM antibody (Monoclonal Anti-EpCAM Antibody, U.K.). Circulating tumor cells (select breast cancer cells MCF-7 as a representative) ( U.S.A) was incubated on sputtered gold foil after binding the antibody for 12 hours. Micrographs of cells on the surface of gold foil such as Figure 4 shown.

[0064] The MCF-7 cells select...

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Abstract

The invention discloses a magnetic bead for capturing circulating tumor cells and a preparation method thereof, and belongs to the technical field of biomedical engineering. The magnetic bead takes anFe3(subscript)O4(subscript) magnetic bead as a core and Au as a shell; and a sulfur group monomolecular layer is modified on the surface of the Au shell, and the carboxyl group on the sulfur group monomolecular layer is activated. The magnetic bead of the invention has good dispersibility, biocompatibility and stability, and has relatively low physiological toxicity. The magnetic bead can be bound with a variety of antibodies, and has high separating and capturing effects after binding with cells.

Description

technical field [0001] The invention belongs to the technical field of biomedical engineering, and in particular relates to a magnetic bead for capturing circulating tumor cells and a preparation method thereof. Background technique [0002] Circulating tumor cells (CTC) refer to all kinds of tumor cells that break away from the primary tumor or metastases of solid tumors, undergo epithelial-mesenchymal transition, and invade peripheral blood. CTCs proliferate in the circulatory system and may form new cancer metastases through adhesion. According to the latest statistical report (Chen Wanqing, Sun Kexin, Zheng Rongshou, et al. Analysis of the incidence and death of malignant tumors in different regions in China in 2014 [J]. Chinese Cancer, 2018 (1)), malignant tumors have become the main cause of death in our country. The detection of CTC can be used for early diagnosis of cancer, monitoring of tumor metastasis, and evaluation of cancer prognosis. The number of CTCs in pe...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0694C12N2509/00
Inventor 任大海勾易行刘佳文尤政
Owner TSINGHUA UNIV
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