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Mussel bionic polypeptide composite magnetic beads and preparation method and application thereof

A biomimetic polypeptide and mussel technology, applied in the field of cell separation and analysis, can solve problems such as difficult release, unfavorable cell re-cultivation analysis research, cell endocytosis, etc., achieve low cost, improve non-specific adsorption resistance, and high cell activity Effect

Active Publication Date: 2019-07-26
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, the magnetic beads used for the enrichment of circulating tumor cells are mostly nano-magnetic beads with a particle size of 10-200nm. There are a large number of magnetic beads on the surface of the enriched and separated cells, which are difficult to release and are easily endocytized by cells. It is not conducive to the re-cultivation and subsequent analysis of captured cells in the later stage. Therefore, it is of great research significance to develop a magnetic bead that can enrich, separate and release circulating tumor cells.

Method used

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  • Mussel bionic polypeptide composite magnetic beads and preparation method and application thereof
  • Mussel bionic polypeptide composite magnetic beads and preparation method and application thereof
  • Mussel bionic polypeptide composite magnetic beads and preparation method and application thereof

Examples

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Embodiment 1

[0041] In this embodiment, fluorescently labeled mussel biomimetic polypeptide composite magnetic beads were prepared through the following steps;

[0042] (1) Preparation of acrylamide-modified magnetic microspheres: take 40 mg of silica-coated Fe 3 o 4 Microspheres (commercially available, particle size 3um) and 500uL 3-(trimethoxysilyl)propyl methacrylate were dispersed in 4.5mL methanol solution, reacted at 37°C for 12h, and applied a magnetic field (116mT) After washing with methanol and distilled water, the acrylamide-modified magnetic microspheres were prepared;

[0043] (2) Preparation of phenylboronic acid-based polymer-modified magnetic microspheres: take 20 mg of acrylamide-based modified magnetic microspheres prepared in step (1), 20 mg of functional monomer CFPBA, 312 mg of hydrophilic monomer HEMA and 2.5 mg of persulfuric acid Ammonium (APS), 0.5uL N,N,N',N'-tetramethylethylenediamine (TEMED) was dissolved in 3mL distilled water to make an aqueous solution, af...

Embodiment 2

[0047] In this embodiment, mussel biomimetic polypeptide composite magnetic beads were prepared through the following steps;

[0048] (1) Preparation of acrylamide-modified magnetic microspheres: take 40 mg of silica-coated Fe 3 o 4 Microspheres (commercially available, particle size 3um) and 250uL 3-(trimethoxysilyl)propyl methacrylate were dispersed in 4.5mL ethanol solution, reacted at 37°C for 24h, and applied a magnetic field (116mT) Acrylamide-modified magnetic microspheres were obtained by washing with methanol and distilled water;

[0049] (2) Preparation of phenylboronic acid-based polymer-modified magnetic microspheres: take 20 mg of acrylamide-based modified magnetic microspheres prepared in step (1), 60 mg of functional monomer AAPBA, 639 mg of hydrophilic monomer HEAA, and 2.5 mg of photoinitiator Reagent 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone (HHMP) in 3mL methanol solution, deoxidized by nitrogen for 30min, UV-induced polymerization for 10min, an...

Embodiment 3

[0053] In this embodiment, mussel biomimetic polypeptide composite magnetic beads were prepared through the following steps;

[0054] (1) Preparation of acrylamide-modified magnetic microspheres: take 40 mg of silica-coated Fe 3 o 4 Microspheres (commercially available, particle size 3um) and 500uL 3-(trimethoxysilyl)propyl methacrylate were dispersed in 4.5mL methanol solution, reacted at 37°C for 18h, and applied a magnetic field (116mT) Wash with methanol and distilled water to obtain acrylamide-modified magnetic microspheres;

[0055] (2) Preparation of phenylboronic acid-based polymer-modified magnetic microspheres: take 20 mg of acrylamide-based modified magnetic microspheres prepared in step (1), 30 mg of functional monomer CFPBA, 213 mg of hydrophilic monomer HPMA and 0.5 mg of persulfuric acid Ammonium, 2.5uL N,N,N',N'-Tetramethylethylenediamine was dissolved in 5mL distilled water to form an aqueous solution. After deoxygenation by nitrogen gas for 20min, the redox...

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Abstract

The invention belongs to the field of cell separation and analysis and particularly relates to mussel bionic polypeptide composite magnetic beads and a preparation method and application thereof. Themussel bionic polypeptide composite magnetic beads utilize the boron affinity reaction of phenylboronic acid and 3,4-dihydroxyphenylalanine to fix a mussel bionic polypeptide with one end containing atumor cell specific recognition sequence to the surface of a magnetic microsphere modified by a phenylboronic acid-based high molecular polymer, and are used for capturing and releasing circulating tumor cells. The mussel bionic polypeptide composite magnetic beads have the advantages of adoption of the simple and easily available raw materials, low cost, simple preparation process, mild reactionconditions, difficult deterioration, degradation and the like of the mussel bionic polypeptide, and have strong practicability.

Description

technical field [0001] The invention belongs to the field of cell separation and analysis, and in particular relates to a mussel biomimetic polypeptide composite magnetic bead and its preparation method and application. Background technique [0002] Circulating tumor cells (CTCs) are tumor cells that exist in the peripheral blood circulation system. Tumor cells in primary or secondary lesions enter the peripheral blood circulation system through spontaneous or diagnostic and therapeutic operations, and are the direct source of tumor metastases. Enrichment and separation of circulating tumor cells, as a liquid biopsy technique, has important clinical significance in the diagnosis, treatment and prognosis monitoring of tumors. Circulating tumor cells were analyzed to elucidate tumor heterogeneity at the level of CTCs. [0003] The content of circulating tumor cells in each milliliter of blood sample is one to dozens. Compared with complex blood samples, the number is extremel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K17/08C12N5/09
CPCC07K17/08C12N5/0693C12N2509/10
Inventor 潘国庆刘磊田小花马悦
Owner JIANGSU UNIV
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