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Magnetic cell and preparation method and application thereof

A cell and cell surface technology, applied in the field of immunoassay, can solve problems such as cumbersome steps, and achieve the effects of simple operation, technical improvement, and simple preparation method

Active Publication Date: 2021-02-26
XUANWU HOSPITAL OF CAPITAL UNIV OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The use of magnetic beads to bind cells is usually applied to cell sorting. Using magnetic bead technology to separate cells, it is necessary to find a certain type of cell-specific surface molecular markers, and then obtain specific antibodies against it, and then couple the antibodies to the magnetic beads , in order to carry out subsequent cell sorting and realize the connection between magnetic beads and cells, the steps are relatively cumbersome
If the magnetic beads can be directly connected with the cells and realize effective application, it must be a more ideal choice and method, but in this field, there is no product and application of the direct combination of magnetic beads and cells

Method used

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  • Magnetic cell and preparation method and application thereof
  • Magnetic cell and preparation method and application thereof
  • Magnetic cell and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069]Example 1 Detection of carboxyl magnetic cells

[0070]Preparation of materials, reagents and instruments:

[0071](1) Magnetic beads: carboxyl magnetic beads, diameter 1μm, 10mg / mL;

[0072](2) Cell line: HEK293T cell;

[0073](3) Magnetic stand: Axygen's IMAG handheld magnetic bead separation device-96P and DYNAMAG-2 magnetic stand;

[0074](4) Mixer: HULAMIXER SAMPLE MIXER

[0075](5) Plasmid: NF155 antigen plasmid and control plasmid (empty vector: pEGFP-N1);

[0076](6) Transfection reagent: PEI Polysciences, make 1mg / ml with distilled water for later use;

[0077](7) Primary antibody: NF155 positive serum, NF155 negative serum;

[0078](8) Buffer: PBS pH 7.4, MES pH 5.0;

[0079](9) Secondary antibody: Goat anti-human IgG (H+L) cross-adsorbed secondary antibody, labeled with Alexa Fluor 555;

[0080]Using the above reagents and consumables, the cell-magnetic bead complex can be obtained by the following method, and the antibody screening can be performed. The sample to be tested in this example has been...

Embodiment 2

[0105]The positive sera of AQP4 antibody and MOG antibody were tested with the same method as in Example 1, and the results were as follows:Figure 4 ,Figure 5 As shown, after the positive serum and the magnetic cells reacted, both showed a red fluorescent signal. Obvious red fluorescent signal can be seen on the cell membrane, while the red fluorescent signal is almost invisible in the negative serum, which is consistent with the actual situation.

Embodiment 3

[0106]Example 3 Detection of Amino Magnetic Cells

[0107]Preparation of magnetic cell detection method:

[0108]Amino magnetic beads: diameter 500nm, 10mg / mL;

[0109]The rest of the reagents, materials and instruments are the same as in Example 1.

[0110]The sample to be tested in this embodiment is a Caspr1 positive and negative serum that has been identified.

[0111]1. Transfection of Caspr1 antigen plasmid and control plasmid:

[0112]Follow the method of "Transfection of NF155 Antigen Plasmid and Control Plasmid" in Example 1.

[0113]2. Magnetic cell preparation:

[0114]Activation of magnetic beads:

[0115]1) After mixing the magnetic beads, take 100μl of amino magnetic beads (10mg / ml) into a 1.5ml centrifuge tube, use DYNAMAG-2 magnetic stand to remove the supernatant, magnetically separate with 1ml PBS pH 8.0 solution, wash 2 Second, remove the supernatant;

[0116]2) Add 100μl of freshly prepared glutaraldehyde solution (15%) to the centrifuge tube containing magnetic beads obtained in step 1), use...

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Abstract

The invention discloses a magnetic cell as well as a preparation method and application thereof. The magnetic cell comprises a magnetic bead and a cell, wherein the magnetic bead is a biological magnetic bead modified by an active functional group, the cell is a cell for overexpression of antigen membrane protein, and the magnetic bead and the cell are coupled together through the active functional group of the magnetic bead and the membrane protein of the cell to form the magnetic cell. The magnetic beads are directly anchored to the cells for the first time, so that the combination of the magnetic beads and the cells is realized without any antigen and antibody reaction; the magnetic frame is used for adsorbing the magnetic cells to realize the control of the cells, so that the operationis simple, the use is convenient, and the automation can be realized by combining with a chemiluminescence microwell plate detection system; besides, when applied to immunodetection, the magnetic cell has high sensitivity and specificity of living cell detection and simplicity in operation of a paramagnetic particle method, and has very important research significance and practical value.

Description

Technical field[0001]The invention relates to the technical field of immunoassays, in particular to a magnetic cell and its preparation method and application.Background technique[0002]Magnetic bead technology is a technical method that appeared in the 1980s. The magnetic beads consist of core metal particles (Fe2O3, Fe3O4), the polymer material (such as polystyrene, polyvinyl chloride) and the outermost functional ligand (such as -NH2, COOH, -OH, -CHO), it is a nano-level superparamagnetic particle. When located in a magnetic field, it will exhibit magnetism and can be captured and absorbed by the magnetic frame. Magnetic bead technology is based on immunology and penetrates into various fields such as physiology, pharmacology, microbiology, biochemistry, and molecular biology. It has become more and more widely used in immunoassays, cell separation, purification of biological macromolecules, and molecular biology. .[0003]At present, magnetic bead technology is widely used in the p...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/543G01N33/564G01N33/58G01N21/76G01N21/64
CPCG01N33/531G01N33/54326G01N33/564G01N33/582G01N21/76G01N21/6458G01N2333/70503G01N2333/47
Inventor 郝峻巍刘亮徐芳王菲
Owner XUANWU HOSPITAL OF CAPITAL UNIV OF MEDICAL SCI
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