Double-targeting magnetic fluorescent nano-microsphere, preparation method thereof and application of double-targeting magnetic fluorescent nano-microsphere in liver cancer circulating tumor cells

A technology of fluorescent nano and nano microspheres, which is applied in the field of biomaterial sensing, can solve the problems of difficult real-time dynamic monitoring, false negative signals, inconsistencies, etc., and achieve improved capture efficiency and detection purity, high-resolution fluorescence imaging, Effect of Precise Dual Targeting Performance

Active Publication Date: 2021-02-19
DALIAN UNIV OF TECH
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Problems solved by technology

However, considering the high recurrence rate and minimal number of HCC-CTCs in the bloodstream, using EpCAM as a single target to isolate CTCs, the detection rate is only between 30% and 80%.
The separation technology based solely on EpCAM mainly has the following disadvantages: (1) The experimental results of simulated CTCs in vitro are inconsistent with the experimental results in vivo, or due to the high heterogeneity and phenotypic changes of CTCs, resulting in missed capture and false negative signals; ( 2) False positive results due to release of benign epithelial cells into circulation due to inflammation or other liver disease
The disadvantages of these studies include: (1) long preprocessing time and low detection efficiency; (2) the morphology of CTCs changes during the extraction process; (3) it is difficult to achieve long-term real-time dynamic monitoring

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  • Double-targeting magnetic fluorescent nano-microsphere, preparation method thereof and application of double-targeting magnetic fluorescent nano-microsphere in liver cancer circulating tumor cells
  • Double-targeting magnetic fluorescent nano-microsphere, preparation method thereof and application of double-targeting magnetic fluorescent nano-microsphere in liver cancer circulating tumor cells
  • Double-targeting magnetic fluorescent nano-microsphere, preparation method thereof and application of double-targeting magnetic fluorescent nano-microsphere in liver cancer circulating tumor cells

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Embodiment Construction

[0047] The specific embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings, but it should be understood that the protection scope of the present invention is not limited by the specific embodiments.

[0048] 1. Preparation method:

[0049] The preparation and response mechanism of the dual-target fluorescent-magnetic nanospheres proposed by the present invention for liver cancer circulating tumor cells are as follows: figure 1 As shown in A–C, there are two steps in the construction of nanosphere MB-MLP-EpCAM. First, MB-PEG was synthesized by a one-pot method using carboxyl-modified ferric oxide (MB-COOH) as the substrate. MLP-PEG was co-assembled with a clickable PEG tetrazine (TZ-PEG) onto MB-COOH via an EDC / DMAP reaction to form MB-PEG, where the ratio of MLP-PEG and TZ-PEG on the modification was 2 :3. Then, through the Diels-Alder-click reaction between tetrazine and dienophile, the TCO-PEG 4 -NHS-EpCAM (TC...

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Abstract

The invention discloses a double-targeting magnetic fluorescent nano-microsphere, a preparation method thereof and an application of the double-targeting magnetic fluorescent nano-microsphere in livercancer circulating tumor cells. According to the nano-microsphere, a carboxyl-modified magnetic ferroferric oxide microsphere is used as a matrix, and anti-epithelial cell adhesion molecules and a fluorescent probe with strong affinity with aminopeptidase are introduced into the surface of magnetic ferroferric oxide at the same time, so that the double-targeting magnetic fluorescent nano-microsphere is obtained. On the basis of no damage of the cell activity (more than 90%), the double-targeting nano-microsphere greatly enhances the capture efficiency (more than 90%) and capture purity (morethan 90%) of the liver cancer circulating tumor cells. More importantly, the nanoparticle has accurate dual-targeting performance, high-resolution fluorescence imaging and excellent selectivity, so that the nano platform becomes a first case to realize real-time recognition imaging and monitoring of in-vivo circulating tumor cells.

Description

technical field [0001] The invention belongs to the technical field of biomaterial sensing, in particular to a dual-target magnetic fluorescent nanometer microsphere, its preparation method and its application in liver cancer circulating tumor cells. Background technique [0002] Liver cancer is a malignant and heterogeneous disease that causes nearly 1 million liver cancer-related deaths each year. High recurrence rate (almost 70% within 5 years after hepatectomy) and distant metastasis are the leading causes of HCC becoming the second leading cancer-related death. Traditional imaging diagnosis and staging assessment of circulating tumor cells in liver cancer have detection limitations, leading to an underestimation of the true extent of the disease. Therefore, in the complex biological research of liver cancer, it is urgent to find effective and representative biomarkers for early diagnosis, recurrence prediction and prognosis evaluation of liver cancer. Metastatic sprea...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/12A61K49/14A61K49/18A61K49/00
CPCA61K49/14A61K49/126A61K49/1866A61K49/186A61K49/0093A61K49/0056
Inventor 邵堃彭孝军夏文汐樊江莉
Owner DALIAN UNIV OF TECH
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