Magnetic red blood cell clusters for enriching circulating tumor cells based on magnetic separation method

A magnetic separation and tumor cell technology, applied in the field of biomedicine, can solve the problems of adverse effects on the purity of CTCs, difficulty in removing white blood cells, and affecting the separation purity of CTCs

Active Publication Date: 2020-10-27
SHENZHEN RES INST OF WUHAN UNIVERISTY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since the physical properties of CTCs and leukocytes overlap to a certain extent, it is difficult to effectively remove leukocytes from isolated CTCs, which makes it difficult to achieve high efficiency and high purity at the same time with a separation method based only on one physical property.
The nanostructures of modified antibodies can specifically target the antigens on CTCs. Although the antigen-antibody binding has high specificity, due to the obvious non-specific adsorption of the nanostructures, it will also adversely affect the purity of the actual isolated CTCs.
[0004] Immunomagnetic nanoparticles are a promising nanostructure for modifying antibodies. After targeting CTCs, CTCs can be efficiently enriched by simple magnetic separation. Adsorption, which greatly affects the separation purity of CTCs
The US FDA-approved commercial CellSearch platform uses immunomagnetic nanoparticles to separate CTCs. However, there is a high background of white blood cells in the obtained CTCs, so the purity is low, only 0.1% to 1.4%.

Method used

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  • Magnetic red blood cell clusters for enriching circulating tumor cells based on magnetic separation method
  • Magnetic red blood cell clusters for enriching circulating tumor cells based on magnetic separation method
  • Magnetic red blood cell clusters for enriching circulating tumor cells based on magnetic separation method

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Embodiment 1

[0053] The features and advantages of the present invention can be further understood through the following detailed description in conjunction with the accompanying drawings. The examples provided are only illustrative of the method of the present invention and do not limit the rest of the present disclosure in any way. [Example 1] Preparation of normal red blood cell clusters using 5-6 μm magnetic microspheres with carboxyl groups on the surface

[0054] Modification process of red blood cells: Mix 30 μL fresh red blood cells from healthy people with 100 μL PBS solution of DSPE-PEG-FA (1 mg / mL) with a molecular weight of 5000, incubate at 4°C for 30 minutes, centrifuge and wash three times with PBS to obtain folic acid Modified red blood cells. Then mix the erythrocytes with surface-modified folic acid and 10 mg / mL polybrene PBS solution, incubate at 4°C for 30 minutes, wash with PBS for three times to obtain erythrocytes with surface-modified polybrene and folic acid, and ...

Embodiment 2

[0056] The preparation process of fluorescent magnetic red blood cell clusters: first mix 30 μL of fresh red blood cells from healthy people and 1000 μL of DiI (10 μg / mL) in PBS solution, incubate at 37 ° C for 60 minutes, and wash three times with PBS to obtain DiI The stained red blood cells were prepared according to the steps described above to obtain DiI-stained magnetic red blood cell clusters. The fluorescent photos taken under a fluorescent microscope are as follows: image 3 (b) shown. [Example 2] Magnetic red blood cell clusters or fluorescent magnetic red blood cell clusters capture MCF-7 cells

[0057] 1. Magnetic erythrocyte clusters capture MCF-7 cells in PBS: Take 50 μL of magnetic erythrocyte cluster suspension (density 1.2×10 7 cells / mL), add 10uL of MCF-7 cell suspension (density 5×10 5 cells / mL), after mixing evenly, place it at 4°C and incubate for 120min, and blow off the mixture at the bottom of the centrifuge tube after incubation. The bright field pho...

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Abstract

The invention discloses magnetic red blood cell clusters for enriching circulating tumor cells based on a magnetic separation method, and belongs to the technical field of biological medicines. Red blood cells are combined with nanometer-grade magnetic microspheres to obtain a red blood cell cluster type biomimetic material, and polybrene is modified on the surfaces of the red blood cells, so thatelectrostatic repulsion originally existing between the red blood cells can be eliminated, and further, the red blood cells can be tightly covered on the surfaces of the microspheres in the total layer. The red blood cells pack the nanometer-grade magnetic microspheres to prevent non-specific adsorption of white blood cells on the surfaces of the microspheres, and besides, folic acid (FA) modified on the surfaces of the red blood cell clusters or an antibody specially recognized or combined with a CTCs surface biomarker can realize direct targeting on CTCs, and finally, in a magnetic separating manner, the white blood cells in blood samples of a clinical patient can be removed, so that high-purity CTCs can be efficiently obtained.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a magnetic red blood cell cluster for enriching circulating tumor cells based on a magnetic separation method. Background technique [0002] Local or regional tumor recurrence and distant metastasis pose a severe challenge to the early diagnosis and cure of tumors. In recent years, molecular biology and clinical studies have shown that circulating tumor cells (circulating tumor cells, CTCs) lead to tumor invasion and the occurrence of micro-metastasis. CTCs are formed by the original tumor cells detaching from the primary tumor and entering the blood system of the human body. After circulating and diffusing in the peripheral blood, CTCs are finally transferred to other tissues and organs of the human body, and then form the same type of tumor as the original tumor Secondary tumors, that is, caused by the metastasis and recurrence of tumors and cancers. By captur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 刘威彭伟
Owner SHENZHEN RES INST OF WUHAN UNIVERISTY
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