Immune nanometer cellulose magnetic liposome and preparation method thereof

A nanocellulose and magnetic liposome technology, applied in the field of biological nanomagnetic materials, can solve the problems of reducing antibody activity, high temperature, and difficulty in maintaining activity, and achieve the effect of good biocompatibility

Active Publication Date: 2014-08-06
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 In the middle of the reaction for 24 hours, the 8000-10000 molecular weight dialysis bag is dialyzed to remove the unreacted condensing agent; the antibody precursor DSPE-PEG-EpCAM is obtained after lyophilization.

[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 solu...

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 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 nanometer magnetic materials used for biology, relates to a nanometer magnetic particle, particularly to an immune nanometer cellulose magnetic liposome and a preparation method thereof. The immune nanometer cellulose magnetic liposome adopts a core-shell structure; the inner core adopts a magnetic material, Fe3O4; a middle layer adopts the mixed material of the liposome and the nanometer cellulose; an outermost layer adopts an antibody wrapping the middle layer; the quality content of the nanometer cellulose is 0.1%-50%; the quality content of the liposome composition is 0.5%-90%; the quality content of the magnetic material is 0.1%-50%. The immune nanometer cellulose magnetic liposome provided by the invention is excellent in biocompatibility; through coupling with the aglucon inside the bioactive substance, the immune nanometer cellulose magnetic liposome can identify and be combined with corresponding antigen, antibody or nucleic acid and the like, so as to rapidly conduct separation and enrichment in an external magnetic field; the immune nanometer cellulose magnetic liposome can be used for detection, separation and purification of biomaterial and has a broad application prospect in biological areas of cell separation, immobilized enzyme, targeted drug, immune detection and the like.

Description

technical field [0001] 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. Background technique [0002] Immunomic microspheres technology (Immunomicrospheres) is a technology based on immunology, using various microspheres coated with immune active substances for immunological or other biological analysis and separation. Since Singer adsorbed human IgG on polystyrene latex particles in 1956 and established the latex agglutination test for detecting rheumatoid factor for the first time, immune microsphere technology has been used in immunology, microbiology, Various fields such as biochemistry, molecular genetics and molecular biology have emerged. [0003] Immunomagnetic Microspheres (IMMS) or Immunomagnetic Microspheres for short, as a new type of functional material, has been widely studied in recent years...

Claims

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Application Information

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Patent Type & Authority Applications(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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