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Core-shell type magnetic composite microsphere for separation and purification of recombinant proteins and preparation method thereof

A technology for separation and purification of magnetic composite microspheres, which is applied in peptide preparation methods, microsphere preparation, chemical instruments and methods, etc., can solve problems such as uneven particle size distribution, lack of surface, and high price, and achieve regular structure, The effect of simple preparation process and low price

Inactive Publication Date: 2014-03-26
SHANGHAI YAOKE BIOTECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, nitrilotriacetic acid ligands have been widely used, and their prices are relatively expensive. At present, the magnetic composite microspheres still have problems such as uneven particle size distribution, low magnetic content, and lack of sufficient active functional groups on the surface.

Method used

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  • Core-shell type magnetic composite microsphere for separation and purification of recombinant proteins and preparation method thereof
  • Core-shell type magnetic composite microsphere for separation and purification of recombinant proteins and preparation method thereof
  • Core-shell type magnetic composite microsphere for separation and purification of recombinant proteins and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Core-shell Fe with a layer thickness of about 50 nm and a cross-linking degree of 75% 3 O 4 / PMG / IDA-Ni 2+ Microspheres

[0040] 1. Preparation of sodium citrate-stabilized magnetic clusters

[0041] 1.3g of ferric chloride hexahydrate (FeCl 3 ·6H 2 O), 3.8 g ammonium acetate (NH 4 Ac), after 0.4g of sodium citrate was dissolved in 70mL of ethylene glycol, it was added to a 150mL three-necked flask, then heated to 160°C, and after the reaction was incubated for 1h, the liquid in the flask was transferred into a 100mL-containing polytetrafluoroethylene lined high-pressure reaction kettle, and then put the reaction kettle into a 200 ℃ oven for 16 hours, take it out, and cool it to room temperature with tap water. The product is separated by magnetic separation, and the unreacted reactant is removed by washing with absolute ethanol, and finally the product is dispersed in absolute ethanol for subsequent use;

[0042] 2. Active vinyl modification on the su...

Embodiment 2

[0050] Example 2: Preparation of core-shell Fe with a shell thickness of about 50 nm and a cross-linking degree of 60% 3 O 4 / PMG / IDA-Ni 2+ Microspheres

[0051] 1. The preparation of sodium citrate-stabilized magnetic clusters is the same as described in Example 1-1;

[0052] 2. The active vinyl modification on the surface of the magnetic cluster is the same as that described in Example 1-2;

[0053] 3. Core-shell Fe 3 O 4 The preparation of / PMG is the same as that described in Examples 1-3, and the dosages of glycidyl methacrylate and 2,2-azobisisobutyronitrile in this example are 200 μL and 10 mg, respectively;

[0054] 4. The ring-opening reaction of iminodiacetic acid is the same as described in Example 1-4;

[0055] 5. Complex Ni 2+ The reactions are as described in Examples 1-5.

Embodiment 3

[0056] Example 3: Preparation of core-shell Fe with a shell thickness of about 50 nm and a cross-linking degree of 50% 3 O 4 / PMG / IDA-Ni 2+ Microspheres (such as figure 1 shown)

[0057] 1. The preparation of sodium citrate-stabilized magnetic clusters is the same as described in Example 1-1;

[0058] 2. The active vinyl modification on the surface of the magnetic cluster is the same as that described in Example 1-2;

[0059] 3. Core-shell Fe 3 O 4 The preparation of / PMG is the same as that described in Examples 1-3; in this example, the dosages of glycidyl methacrylate and 2,2-azobisisobutyronitrile are 300 μL and 12 mg, respectively;

[0060] 4. The ring-opening reaction of iminodiacetic acid is the same as described in Example 1-4;

[0061] 5. Complex Ni 2+ The reactions are as described in Examples 1-5.

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Abstract

The invention belongs to the technical field of nano functional materials, and particularly relates to a core-shell type magnetic composite microsphere for the separation and purification of recombinant proteins and a preparation method thereof. A core of the core-shell type magnetic composite microsphere disclosed by the invention is a magnetic ferroferric oxide nano particle cluster, and a shell of the core-shell type magnetic composite microsphere is a crosslinked polymer network containing epoxy groups. The core-shell type magnetic composite microsphere is prepared through the following method implemented through the steps that: a magnetic nano particle cluster with stable sodium citrate is prepared firstly; then, by using a sol gel method, an active vinyl functional group is modified on the surface of the magnetic cluster; a high-magnetic-responsiveness monodispersed core-shell type magnetic composite microsphere of which the surface is rich in epoxy groups is prepared through distilling-precipitation polymerization; an iminodiacetic acid is adopted for carrying out ring-opening reaction with the epoxy groups and complexing nickel ions; finally, histidine marked proteins are separated and purified. The preparation method of the core-shell type magnetic composite microsphere is simple, controllable in process, and high in efficiency of separation and purification of recombinant proteins. According to the method disclosed by the invention, the thickness and crosslinking degree of a polymer shell layer and the density of surface functional groups can be precisely controlled, therefore, the method has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of nanometer functional materials, relates to magnetic composite microspheres, and in particular relates to a core-shell magnetic composite microsphere for separating and purifying recombinant proteins and a preparation method thereof. Background technique [0002] In recent years, organic-inorganic hybrid composite microspheres, especially magnetic composite microspheres, are receiving extensive attention. Because the magnetic polymer microspheres have both the magnetic responsiveness of inorganic magnetic materials and the surface modifiability of organic polymers, they can easily, quickly and efficiently separate target biomolecules under an external magnetic field. Therefore, it has broad application prospects in biomedical fields such as protein purification, cell separation, magnetic resonance detection and magnetic targeting drug loading. [0003] The prior art discloses that magnetic polymer composi...

Claims

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Application Information

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IPC IPC(8): C08F292/00C08F8/44C08F8/32H01F1/26B01J13/02C07K1/22
Inventor 汪长春
Owner SHANGHAI YAOKE BIOTECH DEV
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