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Preparation method and applications of reduction self-assembled protein-coated magnetic microspheres

A magnetic microsphere and protein technology, applied in the field of materials, can solve the problems of high non-specific adsorption, uneven product particle size, and low non-specific adsorption, so as to achieve difficult aggregation, avoid aldehyde residues, and easy particle size The effect of regulation

Active Publication Date: 2017-07-28
HUAZHONG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The purpose of the present invention is to overcome the defects of uneven product particle size, high non-specific adsorption, poor stability and complicated preparation process in the preparation of protein-coated magnetic microspheres in the prior art, and to provide a method with simple preparation process, low cost, Preparation method and application of water-soluble magnetic microspheres with advantages of good stability, high biocompatibility and low non-specific adsorption

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  • Preparation method and applications of reduction self-assembled protein-coated magnetic microspheres
  • Preparation method and applications of reduction self-assembled protein-coated magnetic microspheres
  • Preparation method and applications of reduction self-assembled protein-coated magnetic microspheres

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Embodiment 1

[0046] A preparation method for reducing self-assembled protein-wrapped magnetic microspheres, comprising the following steps:

[0047] (1) Physical mixing: take a certain volume of superparamagnetic iron oxide nanoparticles (SPIO), and ensure that the mass of SPIO is 5 mg. After adding 3 mL of ethanol, vortex for 30 s, collect SPIO with a magnet, remove the supernatant, add 100 μL of n-hexane to re-dissolve SPIO, then add 100 μL of ethanol, 1 mL of pH 7.2 7 Tris buffer. Sonicate in an ultrasonic cleaner for 3min, transfer SPIO to a 25mL glass bottle and continue to sonicate for 1min, then add 4mL of pH 7.27 Tris buffer while sonicating, and then sonicate for 2min, add 50mg of bovine serum albumin (BSA) into SPIO, and mix the plasma Mix on a homogenizer for 12 hours to obtain a coarse emulsion.

[0048] (2) Reduction of self-assembled package: Transfer the crude emulsion to a 25mL round bottom flask, add 1.38mol / L 3-mercapto-1,2-propanediol in a 37°C water bath with mechanica...

Embodiment 2

[0058] A preparation method for reducing self-assembled protein-wrapped magnetic microspheres, comprising the following steps:

[0059] (1) Physical mixing: take a certain volume of superparamagnetic iron oxide nanoparticles (SPIO), and ensure that the mass of SPIO is 5 mg. After adding 3 mL of ethanol, vortex for 30 s, collect SPIO with a magnet, remove the supernatant, add 100 μL of chloroform to re-dissolve SPIO, then add 100 μL of ethanol, 1 mL of pH 7.27 Tris buffer. Sonicate in an ultrasonic cleaner for 3min, transfer SPIO to a 25mL glass bottle and continue to sonicate for 1min, then add 4mL pH 7.27 Tris buffer while sonicating, and then sonicate for 2min, add 50mg of BSA to SPIO, and mix on a plasma mixer for 12h A crude emulsion is obtained.

[0060] (2) Reduction of self-assembled package: Transfer the crude emulsion to a 25 mL round bottom flask, add 1.838 mol / L mercaptoethanol under 37°C water bath and 300 rpm mechanical stirring, and react for 6 min.

[0061] (3...

Embodiment 3

[0066] A preparation method for reducing self-assembled protein-wrapped magnetic microspheres, comprising the following steps:

[0067] (1) Physical mixing: take a certain volume of superparamagnetic iron oxide nanoparticles (SPIO), and ensure that the mass of SPIO is 5 mg. After adding 3 mL of ethanol, vortex for 30 seconds, collect SPIO with a magnet, remove the supernatant, add 100 μL of n-hexane to re-dissolve SPIO, then add 100 μL of ethanol, 1 mL of pH 7.27 Tris buffer. Sonicate in an ultrasonic cleaner for 3min, transfer SPIO to a 25mL glass bottle and continue to sonicate for 1min, then add 4mL of pH 7.27 Tris buffer while sonicating, and then sonicate for 2min, add 50mg of human serum albumin (HSA) into SPIO, and the plasma mixes Mix on a homogenizer for 12 hours to obtain a coarse emulsion.

[0068] (2) Reduction of self-assembled package: Transfer the crude emulsion to a 25mL round bottom flask, add 1.38mol / L 3-mercapto-1,2-propanediol in a 37°C water bath with mec...

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Abstract

The invention discloses a preparation method of reduction self-assembled protein-coated magnetic microspheres. According to the preparation method, mercaptoethanol, dithiothreitol, or 3-mercapto-1,2-propanediol are taken as reductive agents; reduction reaction is adopted to break the disulfide bonds of protein, and expose the sulfydryl and hydrophobic regions of protein, self-assembling with magnetic nano particles via sulfydryl coordination and hydrophobic interaction is realized so as to obtain protein-coated magnetic microspheres of a core-shell structure. The particle size of the protein-coated magnetic microspheres is convenient to adjust; stability is high; water solubility is high; aggregation is not easily caused; nonspecific adsorption is low; excellent biocompatibility is achieved; no special equipment is needed; consumed time is short; operation is convenient; production cost is low; preparation conditions are mild; direct fixing of specific antibodies in preparation process is realized; the activity of the protein-coated magnetic microspheres coupled with antibodies is high; and the obtained reduction self-assembled protein-coated magnetic microspheres can be used in magnetic immunity chromatography preferably.

Description

technical field [0001] The invention belongs to the field of materials, and more specifically relates to a preparation method of reduced self-assembled protein-wrapped magnetic microspheres and an application of magnetic immunoassay. Background technique [0002] Superparamagnetic iron oxide nanoparticles (SPIO) are inorganic ferromagnetic substances with a particle size between 1 and 100nm. Because of their unique characteristics such as high magnetic responsiveness, superparamagnetism, large specific surface area, and easy separation, It has been widely used in biomedical fields, such as biosensors, cell sorting, tissue engineering, magnetic resonance imaging (MRI), optical multi-channel imaging, targeted drug delivery, hyperthermic treatment of cancer, and immune analysis. [0003] The synthesis methods of SPIO mainly include chemical co-precipitation method, thermal decomposition method, emulsion synthesis method, hydrothermal synthesis method, etc. The naked and unmodif...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/543G01N33/532
CPCG01N33/532G01N33/54326G01N33/558G01N33/577
Inventor 杨海汪业红杨勇张德军栗静杨祥良
Owner HUAZHONG UNIV OF SCI & TECH
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