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Enriching method of circulating tumor cells

A tumor cell enrichment technology, applied in the field of nanomaterials, can solve the problems of cumbersome operation, slow magnetic response, inconvenient for ordinary users, etc., and achieve good enrichment effect, good reproducibility, fast and efficient capture and enrichment Effect

Inactive Publication Date: 2014-03-12
WUHAN UNIV
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Problems solved by technology

This method introduces many steps for the capture of CTCs, and the operation is cumbersome.
However, the MACS system uses 50 nm magnetic nanospheres, which have a slow magnetic response and require strong magnetic separation tools——magnetic separation columns (MACS Columns and MACS Separator) to complete the separation, which is inconvenient for ordinary users

Method used

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  • Enriching method of circulating tumor cells
  • Enriching method of circulating tumor cells
  • Enriching method of circulating tumor cells

Examples

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Embodiment 1

[0024] Example 1 Enrichment and Analysis of Circulating Tumor Cells

[0025] Taking the breast cancer cell SK-BR-3 as an example, the efficient and rapid capture and sensitive detection of circulating tumor cells using magnetic nanospheres will be described in detail below.

[0026] 1. Method

[0027] 1. Preparation of Magnetic Nanospheres

[0028] The preparation of magnetic nanospheres is based on the preparation of fluorescent-magnetic double-coded spheres in our laboratory, that is, using the assembly method to assemble magnetic nanoparticles nano-γ-Fe on the surface of styrene-propionamide nanospheres 2 o 3. Specifically: ① The styrene-acrylamide copolymerized nanospheres were prepared by emulsion-free polymerization, and carboxyl groups were modified on the surface of the nanospheres by hydrazide and succinic anhydride, and the obtained nanospheres were activated by EDC / NHS A layer of polyethyleneimine (PEI) was assembled on the surface of the carboxyl nanospheres,...

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Abstract

The invention provides an enriching method of circulating tumor cells (CTCs) and a kit thereof. The enriching method is applied to quickly and efficiently enriching CTCs from blood. The enriching method comprises the following steps of coupling an anti-epithelial cell adhesion molecule (EpCAM) antibody onto a magnetic nanosphere to prepare an immune-magnetic bead; adding the immune-magnetic bead into a blood sample for hatching for 5 minutes, and realizing separating, enriching and purifying of CTCs by the steps such as magnetic separating and washing. The enriching method disclosed by the invention is high in capturing efficiency, short in enriching time, good in specificity and very reliable. Activity ratio of the captured cell is kept at (90.5 + / -1.2)%, and the captured cell can be directly used for cultivation, reverse transcriptase polymerase chain reaction (RT-PCR) and immune cell chemical analysis. Besides, the enriching method provides powerful means for detecting and researching CTCs, and has extremely important medical significance.

Description

technical field [0001] The invention relates to a method for enriching circulating tumor cells and belongs to the technical field of nanomaterials. Background technique [0002] Cancer is the greatest threat to human health and life in the world today, so that everyone "talks about cancer and turns pale". Malignant tumors of epithelial origin are the most common cancers, including breast cancer, lung cancer, liver cancer, gastric cancer, pancreatic cancer, esophageal cancer, etc. Caused. Studies in recent years have shown that cancer cells can shed from the primary tumor tissue into the blood system and become circulating tumor cells (CTCs), which is an important intermediate event of cancer metastasis. The number of CTCs has a great relationship with the survival rate of cancer patients, so detection CTCs are of great significance in medicine. However, CTCs exist in blood at extremely low concentrations, so the biggest difficulty in detecting CTCs is the low concentratio...

Claims

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Application Information

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IPC IPC(8): G01N1/40
Inventor 庞代文温聪颖吴玲玲张志凌
Owner WUHAN UNIV
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