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Bionic immune magnetic nanoparticles as well as preparation method and application thereof

A technology of magnetic nanoparticles and bionic immunity, applied in the field of biomedicine, can solve problems such as low encapsulation efficiency, complicated procedures, and poor stability, and achieve high capture efficiency, reduce interference, and improve purity

Pending Publication Date: 2022-05-13
SHENZHEN BAY LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, its stability is not good, the program is complicated, and the encapsulation rate is not high

Method used

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  • Bionic immune magnetic nanoparticles as well as preparation method and application thereof
  • Bionic immune magnetic nanoparticles as well as preparation method and application thereof
  • Bionic immune magnetic nanoparticles as well as preparation method and application thereof

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preparation example Construction

[0071] According to a typical embodiment of the present application, a method for preparing the above-mentioned biomimetic immunomagnetic nanoparticles is provided. The preparation method comprises the following steps: S1, preparing neutrophil membrane vesicles; S2, coating magnetic nanoparticles with neutrophil membrane vesicles to obtain biomimetic magnetic nanoparticles. Correspondingly, the preparation method of the biomimetic immune magnetic nanoparticles used for separation includes the above preparation method of the biomimetic immune magnetic nanoparticles and S3, the CTC targeting antibody is physically connected and / or covalently connected to the biomimetic magnetic nanoparticles, the physical connection And / or the covalent linkage is a conjugate linkage or a chemical click linkage.

[0072] In a typical embodiment of the present application, the CTC-targeting antibody is an anti-epithelial cell adhesion molecule antibody.

[0073] In an embodiment of the present ap...

Embodiment approach

[0079] According to a typical embodiment of the present application, S3 includes: treating the biomimetic magnetic nanoparticles prepared in S2 with carboxyl-polyethylene glycol-phospholipid, then adding NHS / EDC for activation, and then cross-linking streptavidin, biological The biomimetic immune magnetic nanoparticles were obtained by binding the primed CTC targeting antibody.

[0080] In another typical embodiment of the present application, S4 is also included, using liquid chromatography tandem mass spectrometry to verify the reduction of protein corona adsorption of Neu-IMNs and Sanger sequencing to identify circulating tumor cells isolated from Neu-IMNs.

[0081] According to a typical embodiment of the present application, an application of biomimetic immunomagnetic nanoparticles for separating CTCs in the preparation of products for enriching, separating or detecting circulating tumor cells is provided.

[0082] According to a typical embodiment of the present applicat...

Embodiment 1

[0085] The preparation and application steps of neutrophil membrane vesicle-coated immunomagnetic nanoparticles (Neu-IMNs) are as follows: (1) Preparation of microfluidic photoporation chip; (2) Preparation of isolated neutrophils from clinical peripheral blood Neu-vesicles; (3) using prepared Neu-vesicles to coat magnetic nanoparticles (MNs) to obtain biomimetic magnetic nanoparticles (Neu-MNs); and (4) combining anti-epithelial cell adhesion molecule (anti-EpCAM) antibody with Neu-MNs are bioconjugated. (5) The reduction of protein corona adsorption of Neu-IMNs was verified by liquid chromatography tandem mass spectrometry, and the circulating tumor cells isolated from Neu-IMNs were identified by Sanger sequencing.

[0086] Specific steps are as follows:

[0087] (1) Preparation of microfluidic photoporation chip:

[0088] Coating: Put the cleaned silicon wafer on the coating machine, spin coat a layer of SU-8-2050 photoresist, turn it forward at 600rpm, run for 30s, then ...

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Abstract

The invention discloses bionic immune magnetic nanoparticles as well as a preparation method and application thereof. The bionic immune magnetic nanoparticles comprise magnetic nanoparticles and neutrophile granulocyte membrane vesicles, and the magnetic nanoparticles are coated with the neutrophile granulocyte membrane vesicles to form the bionic magnetic nanoparticles. The bionic magnetic nanoparticles are subsequently connected with a targeting antibody and can be used for separating biological components. The bionic immune magnetic nanoparticles at least also have the following advantages that the adsorption of non-specific proteins is reduced and the targeting capability of the IMNs in blood is maintained by virtue of the lipid bilayer of Neu-vesicles; the interaction between the CTCs and the neutrophils is enhanced, and the efficiency of separating the CTCs by the IMNs is improved; the interaction between leukocytes and neutrophils is reduced, the interference of WBCs in the separation process is reduced, and the purity of separated CTC is improved; the soft interaction between the Neu-vemics and the CTC increases the vitality of the isolated CTC.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a bionic immune magnetic nanoparticle, its preparation method and application. Background technique [0002] Isolation of circulating tumor cells (CTCs) from clinical peripheral blood samples is a non-invasive liquid biopsy, which has broad prospects for early detection and diagnosis of cancer. So far, a variety of emerging methods have been explored to isolate CTCs from peripheral blood, such as enrichment methods based on physical properties, enrichment methods based on biological characteristics, and enrichment methods without differences in nucleated cells, etc. Among them, the most widely used strategy is based on immunomagnetic nanoparticles (IMNs), in which magnetic nanoparticles are used as separation media and specific antibodies are used as capture ligands. Due to the scarcity of CTCs in peripheral blood (only a few to dozens per milliliter of blood), how to effici...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/551G01N33/554G01N33/531G01N33/569G01N33/574C12N5/09G01N1/40G01N15/10A61K47/68A61K47/69A61K47/02A61P35/00B82Y5/00
CPCG01N33/54346G01N33/5434G01N33/531G01N33/56966G01N33/57492C12N5/0693G01N1/405G01N15/10A61K47/6898A61K47/6925A61K47/6901A61K47/6843A61K47/02A61P35/00B82Y5/00C12N2509/00G01N2015/1006
Inventor 饶浪吴显佳
Owner SHENZHEN BAY LAB
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