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Nano magnetic bead for capturing circulating tumor cells as well as preparation method and application thereof

A technology of tumor cells and nano-magnetic beads, applied in the field of molecular biology, can solve problems such as unapproved, and achieve the effects of reducing interference, reducing non-specific adhesion, and good cell compatibility

Pending Publication Date: 2021-02-02
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many methods have been used to isolate CTCs, such as nano-substrate capture method modified by affinity molecules, microfluidic capture method, density gradient separation method, immunomagnetic bead separation method, etc., but these methods are still not approved for routine clinical detection CTCs

Method used

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  • Nano magnetic bead for capturing circulating tumor cells as well as preparation method and application thereof
  • Nano magnetic bead for capturing circulating tumor cells as well as preparation method and application thereof
  • Nano magnetic bead for capturing circulating tumor cells as well as preparation method and application thereof

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preparation example Construction

[0055] The present invention provides a preparation method and application of nano-magnetic beads coated with hydrogel magnetic beads for capturing circulating tumor cells (CTCs), characterized in that the nano-magnetic beads coated with hydrogel magnetic beads include:

[0056] (1) Synthesize nano-scale magnetic beads (such as 100nm) through a simple hydrothermal synthesis method;

[0057] (2) The surface of the magnetic beads is modified by silanization to double bonds, and the anti-adhesion hydrogel layer is further modified by reflux precipitation polymerization method;

[0058] (3) Introducing affinity molecules such as antibodies with specific capture CTCs on the surface of the hydrogel layer through the condensation reaction of amino groups and carboxyl groups.

[0059] The step (1) includes: synthesizing magnetic beads with a diameter of about 100 nm and good magnetic properties through a simple hydrothermal synthesis method.

[0060] Step (1) specifically comprises:...

Embodiment 1

[0095] The preparation method of the nano-magnetic beads of the hydrogel-coated magnetic beads is as follows:

[0096] a) Iron trichloride hexahydrate (3-6mmol), trisodium citrate (2-4mmol) and 0.5g PEG are dissolved in 70-100ml of ethylene glycol, then add sodium acetate and stir at room temperature, and finally transfer to Reactor, react at 200°C for 6-10h. After the reaction was finished, it was cooled to room temperature, and the product was separated and collected by an external magnetic field and washed with ethanol and deionized water. Store in a refrigerator at 4°C.

[0097] b) The Fe of about 100nm obtained in step a) 3 o 4 Disperse the magnetic nanoparticles in 10-100mL ethanol, add 1-20mL deionized water, 0.1-5mL ammonia water and 0.1-1g MPS, stir, react overnight, then collect the product with the help of a magnet, and wash it with a large amount of ethanol, Disperse in a certain volume of acetonitrile solution, ultrasonically disperse, add SBMA and MAA with a ...

Embodiment 2

[0101] Since the surface of the magnetic beads is modified with EpCAM antibody, we chose K562 cells (EpCAM-) and MCF-7 cells (EpCAM+) to investigate the effect of incubation time on capture efficiency. Since K562 cells are human chronic myeloid leukemia cancer cell lines, they grow in suspension and do not need to be treated with trypsin. A certain amount of cells are directly taken from the culture flask, counted, and finally the cells are prepared to a density of 2x 10 5 / mL of cell solution. MCF-7 cells grow adherently, and are separated from the culture dish by trypsin treatment, suspended in PBS buffer solution, counted, and finally the cells are also prepared into 2x 10 5 / mL density. Take 0.5mL of the above-prepared MCF-7 or K562 cell solution and put it into a 2mL EP tube, then add 0.5mL of PBS solution containing 0.1mg of antibody-modified nano-magnetic beads to the tube, mix well, and put it in the cell culture incubator Incubate for 5-30min. Then, the captured ce...

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Abstract

The invention discloses a nano magnetic bead for capturing circulating tumor cells as well as a preparation method and an application of the nano magnetic bead. The nano magnetic bead is of a core-shell structure and comprises a magnetic bead serving as a core and a hydrogel layer serving as a shell, the surface of the magnetic bead is connected with the hydrogel layer, and the outer surface of the hydrogel layer is modified with capture molecules used for capturing circulating tumor cells. According to the nano magnetic bead, the synergistic effect of the hydrogel layer formed by anti-adhesion molecules and affinity molecules is utilized, so that the non-specific adhesion of blood cells can be reduced to a great extent, and CTCs can be efficiently captured.

Description

technical field [0001] The present invention relates to an anti-adhesion hydrogel magnetic bead, in particular to a nano-magnetic bead for capturing circulating tumor cells and its preparation method and application, such as the application in the separation and identification of circulating tumor cells, belonging to molecular field of biology. Background technique [0002] Circulating tumor cells (CTCs) refer to malignant tumor cells that have shed from the primary tumor or metastatic site into the peripheral blood of the human body. The study of CTCs is expected to clarify the internal mechanism of cancer metastasis, drug sensitivity and drug resistance, so as to realize individualized and effective treatment for cancer patients. However, the number of CTCs in peripheral blood is extremely rare, making it difficult to isolate. Molecular identification and functional analysis of CTCs require high-purity CTCs samples. Many methods have been used to isolate CTCs, such as n...

Claims

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Application Information

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IPC IPC(8): C12N5/09C08F292/00C08F220/38C08F220/06
CPCC12N5/0693C08F292/00C12N2509/00C08F220/06
Inventor 裴仁军王志利
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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