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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 biochemical characteristics of the cells themselves, which is not conducive to the detection of subsequent cells.

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

Embodiment 1

[0043] The preparation of embodiment 1 magnetic beads

[0044] according to figure 1 with figure 2 The flow shown is to prepare the magnetic beads of the present invention.

[0045] Silylated Fe 3 o 4 The magnetic beads are prepared by a conventional method, and their modifiers can be 3-aminopropyltriethoxysilane (APTMS) or 3-mercaptopropyltrimethoxysilane (MPTMS).

[0046] Colloidal gold is prepared by oxidation-reduction method, and its preparation method can be:

[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. Dissolve sodium borohydride in trisodium citrate solution, add colloidal gold pre-solution, and stir rapidly with a glass rod for 5 minutes to obtain colloidal gold. The diameter of the prepared colloidal gold particles is 20nm-30nm.

[0048] The preparation method of magnetic beads comprises the following:

[0049] 1. Preparation of ...

Embodiment 2

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

Embodiment 3

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

[0063] Configure MCH and MUA absolute ethanol solution in the same ratio as the magnetic bead modification in Example 1 in a circular petri dish, soak the coverslip sputtered with gold foil overnight, and then in the PBS solution of pH 7.4 according to the example 1 NHS / EDC weight ratio for NHS / EDC coupling reaction and binding anti-EpCAM antibody (Monoclonal Anti-EpCAM Antibody, U.K.). In the culture medium, the circulating tumor cells (breast cancer cell MCF-7 is selected as a representative) ( U.S.A) Incubated for 12 hours on sputtered gold foil after antibody binding. Micrographs of cells on the surface of gold foil as Figure 4 shown.

[0064] The MCF-7 cells used in this experiment are induced b...

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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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