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A kind of immune nanocellulose magnetic liposome and preparation method thereof

A technology of nanocellulose and magnetic liposome, which is applied in the field of biological nanomagnetic materials, can solve the problems of reducing antibody activity, prone to caking, and difficult to maintain activity, and achieves the effect of good biocompatibility

Active Publication Date: 2016-01-20
上海裕隆医学检验所股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are still many problems in the existing immunomagnetic microspheres, such as lecithin / cholesterol magnetic liposomes, immune sodium alginate magnetic liposomes, PLGA magnetic liposomes, etc.
First of all, the antibody grafting rate in immunomagnetic microspheres is low and the activity is difficult to maintain
The main reason is that the active groups existing on the surface of the magnetic microspheres are used for modification, and the temperature is high and the time is long, which easily reduces the activity of the antibody.
Thirdly, the prepared immunomagnetic microspheres generally have insufficient stability and are prone to agglomeration; after half a year of storage, the samples are prone to agglomeration

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of immune nano-cellulose magnetic liposomes using antibody precursors

[0026] (1) Put 5.0mg DSPE-PEG-NH 2 Mix with 1.0 mg of anti-EpCAM monoclonal antibody, and add 5.0 mL of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCL) at a concentration of 1.0 mg / mL After 24 hours of neutralization, the 8000-10000 molecular weight dialysis bag was dialyzed to remove the unreacted condensing agent; after lyophilization, the antibody precursor DSPE-PEG-EpCAM was obtained.

[0027] (2) Prepare immune nano-cellulose magnetic liposomes by film dispersion method. Mix 10.0 mg of lecithin, 6.0 mg of cholesterol, 2.0 mg of carboxymethyl chitosan stearyl quaternary ammonium salt and 3.0 mg of oil-soluble magnetic nanoparticles in 10.0 mL of oil phase chloroform, incubate for more than 20 minutes, and put In the eggplant-shaped bottle, the lipid film is formed by rotary evaporation;

[0028] (3) Mix the formed lipid membrane with an aqueous solution con...

Embodiment 2

[0029] Example 2 Preparation of Nano-cellulose Magnetic Liposome

[0030] Add lecithin 5.0mg, cholesterol 2.0mg, DSPE-PEG-NH 2 1.0 mg and 1.0 mg of oil-soluble magnetic nanoparticles were dissolved in dichloromethane; mixed as an organic phase, removed the organic solvent to form a film, and then added octadecyl quaternary ammonium salt containing carboxymethyl chitosan (concentration 0.3 mg / mL) and 5.0 mL of carboxymethyl cellulose (nanocellulose concentration 0.5 mg / mL) to hydrate the lipid membrane. The aqueous phase is 0.9% saline or a buffer solution with pH=5 to 9, Ultrasound for more than 30 minutes during hydration, and then magnetically separate more than 3 times for use.

Embodiment 3

[0031] Example 3 Preparation of immunological nano-cellulose magnetic liposomes

[0032] Take the 20% solid content and the 50~100nm particle size of the aminoated nanocellulose magnetic liposome (lecithin / cholesterol / nanocellulose / DSPE-PEG-NH 2 ) Mix 1.0 mL of the solution with 1 mL of the solution containing 2.0 mg of anti-human epidermal growth factor receptor 2 (HER2) monoclonal antibody, and add 5 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide The hydrochloride salt, mixed uniformly, stirred at room temperature for 48 hours, after magnetic separation and washing, the weight percentage of the magnetic beads and the monoclonal antibody contained in the prepared immunomagnetic liposome is 25:1, which can be placed in Store in a mixed solution of water and ethanol (20% by weight of ethanol). This embodiment can be extended to amino or carboxylated magnetic beads with a solid content of 0.5%-30%.

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Abstract

The invention belongs to the technical field of nano-magnetic materials for biology, and relates to a nano-magnetic particle, in particular to an immune nano-cellulose magnetic liposome and a preparation method thereof. It is a core-shell structure, the core is magnetic material Fe3O4, the middle layer is a mixture of liposome and nanocellulose, and the outermost layer is the antibody coated on the middle layer; the mass content of nanocellulose is 0.1%~ 50.0%, the mass content of liposome components is 0.5% ~ 90.0%, and the mass content of magnetic materials is 0.1% ~ 50.0%. The immune nanocellulose magnetic liposome of the present invention has good biocompatibility. The ligand coupling in the substance can recognize and bind to the corresponding antigen, antibody or nucleic acid, etc., and can be separated and enriched rapidly in an external magnetic field. It can be used for the detection and separation and purification of biological materials, and can be used in cell separation, immobilized enzymes, targeted drugs and Immunoassays and other biological fields have broad application prospects.

Description

Technical field [0001] The present invention is a technical field of biological nanomagnetic materials. It involves a nanomagnetic particles, which is specifically for an immunohicic fibrin magnetic lipid and its preparation methods. Background technique [0002] Immunomicrospheres is a technology that is used for immunology or other biological analysis separations based on immunomotive activity based on immunomotive activity.Since Singer adsorbed the human LGG to the styrene gel grain particles in 1956, the immune micro -ball technology has been established for the first time to detect the glue factor in rheumatoid factors for the first time.Biochemistry, molecular genetics and molecular biology are emerging. [0003] Immig magnetic micro -balls (IMMS) or immunomagnetic micro beads for the abbreviation of immunomagnetic micro -beads have been widely studied in recent years. The application in the fields of biomedical and clinical diagnosis has aroused the heights of researchers ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/395A61K47/48A61K47/38A61K47/02A61K47/36G01N33/53C12N5/09
Inventor 穆海东汪宁梅梁晓飞
Owner 上海裕隆医学检验所股份有限公司
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