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Water soluble microsphere preparation method and application thereof

A water-soluble, microsphere technology, applied in the field of materials, can solve the problems of high non-specific adsorption, high toxicity, inactivation of biologically active macromolecules, etc., and achieve good stability.

Active Publication Date: 2016-11-16
武汉纳达康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] In summary, in the prior art, water-soluble microspheres wrapped with biomacromolecules are prepared by emulsification solvent volatilization method, emulsification solidification method and desolvation method, all of which need to increase the stability of biomacromolecules through heat treatment or with the help of chemical cross-linking agents. There are following defectives: the one, adopt cross-linking agent to take place chemical cross-linking, chemical cross-linking is non-specific, and any nucleophilic group (as amine and hydroxyl) existing in the biomacromolecular substance structure has reactivity, causes non-specific The heterosexual adsorption is high; the second is that the thermal denaturation method will irreversibly change the structure of biomacromolecules, resulting in the loss of biological characteristics of biomacromolecules; , glutaraldehyde and other aldehydes will cause the inactivation of biologically active macromolecules

Method used

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  • Water soluble microsphere preparation method and application thereof
  • Water soluble microsphere preparation method and application thereof
  • Water soluble microsphere preparation method and application thereof

Examples

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preparation example Construction

[0042] A preparation method of water-soluble microspheres, comprising the steps of:

[0043] (1) Preparation of coarse emulsion: dissolving oil-soluble nanoparticles in an organic solvent to obtain an oil phase, dissolving water-soluble biomacromolecules containing mercapto groups or disulfide bonds into water to obtain a water phase, and pre-mixing the oil-water two-phase, A crude emulsion is obtained.

[0044] Wherein, the nanoparticles can be oil-soluble superparamagnetic nanoparticles, such as ferric oxide nanoparticles, ferric oxide nanoparticles or other doped iron oxide nanoparticles; the organic solvent is chloroform, dichloromethane, formic acid Ethyl ester, ethyl acetate, n-hexane, cyclohexane, or their mixtures, while containing absolute ethanol as a co-emulsifier; biomacromolecules contain immunoglobulin, albumin, hemoglobin, casein, collagen, Mixing of one or more kinds of enzymes, such as serum, ascites, milk and other biological fluids; the method of premixing ...

Embodiment 1

[0055] A method for preparing water-soluble magnetic microspheres wrapped by biomacromolecules, comprising the following steps:

[0056] (1) Preparation of coarse emulsion: 0.6 mL of chloroform dissolved with 140 mg / mL of superparamagnetic iron oxide nanoparticles, 0.06 mL of absolute ethanol, and 30 mL of an aqueous solution of 4.4 g / L bovine serum albumin were mixed evenly, and superparamagnetic The mass ratio of iron oxide to bovine serum albumin is 1:1.57, and the volume ratio of oil-water two-phase is 1:44.

[0057] (2) Cavitation cross-linking reaction: use a high-shear dispersing emulsifier (FLUKO FA25, Shanghai Fluke Fluid Machinery Manufacturing Co., Ltd.) to process the coarse emulsion prepared in step (1) through 10000 rpm high-shear for 30s, and then In a high-pressure homogenizer (JNBIOJN-02HC, Guangzhou Juneng Biotechnology Co., Ltd.), a uniform microemulsion was formed after 600 bar high-pressure homogenization once.

[0058] (3) Separation and purification: Fi...

Embodiment 2-14

[0062] A method for preparing water-soluble magnetic microspheres wrapped by biomacromolecules, preparing water-soluble magnetic microspheres wrapped by biomacromolecules according to the experimental conditions of Examples 2-14 in Table 1, comprising the following steps:

[0063] (1) Preparation of coarse emulsion: 0.6-4.1 mL of chloroform dissolved with 16.7-180 mg / mL superparamagnetic iron oxide nanoparticles, 0.06-0.41 mL of absolute ethanol, and an aqueous solution of 0.5-250 g / L bovine serum albumin Mix 30mL, the mass ratio of superparamagnetic iron oxide to bovine serum albumin is 1:1.25-20, and the volume ratio of oil-water two-phase is 1:5-200.

[0064] (2) Cavitation cross-linking reaction: use a high-shear dispersing emulsifier (FLUKO FA25, Shanghai Fluke Fluid Machinery Manufacturing Co., Ltd.) to process the coarse emulsion obtained in step (1) through 10000-16000 revolutions per minute high-shear 30-90s, and then in a high-pressure homogenizer (JNBIO JN-02HC, Gua...

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Abstract

The invention discloses a preparation method of water soluble microspheres. Oil solubility nano-particles are dissolved into an organic solvent to obtain an oil phase, water-soluble hydrosulphonyl containing or disulfide bond biomacromolecules are dissolved into water to obtain a water phase; the oil phase and the water phase are premixed, and a crude emulsion is obtained; the crude emulsion is subjected to the cavatition through the physical stimulation means, biomacromolecule is subjected to a crosslinking reaction, and the biological macromolecule shells wrap the surfaces of the nano-particles. The preparation method is simple and mild in reaction conditions, the obtained microspheres have the good water solubility, the grain size is controllable, the grain size is uniform, the stability is good, the non-specific adsorption is low, the biological compatibility is good, the coupling marking is easy, and the method can be widely used for the fields of bioanalysis, environment analysis, food safety testing, sample separation and purification, medical examination, imaging, biological medicine research and the like.

Description

technical field [0001] The invention belongs to the field of materials, and more specifically relates to a preparation method and application of water-soluble microspheres. Background technique [0002] Due to its good magnetic responsiveness, superparamagnetic iron oxide can be easily magnetically separated and guided under the action of an external magnetic field. Since it is not permanently magnetized in a magnetic field, it is safe and easy to control. It can be combined with a variety of biological functional molecules , such as enzymes, DNA, proteins, etc., have broad application prospects in DNA separation and purification, magnetically targeted drug delivery, medical testing, diagnosis, gene therapy, cell separation, immune analysis, immobilized enzymes and proteins, etc. [0003] Nanoparticles prepared by thermal decomposition have small particle size, monodispersity, narrow particle size distribution, high crystallinity and good magnetic properties. It is currently...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/576
CPCG01N33/54326G01N33/5761
Inventor 杨海杨勇梁培培汪业红杨祥良
Owner 武汉纳达康生物科技有限公司
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