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Surface zwitterionic polymer modified magnetic microsphere as well as preparation method and application of magnetic microsphere

A technology of magnetic microspheres and zwitterions, which is applied in the fields of alkali metal compounds, chemical instruments and methods, and other chemical processes, and can solve the problems of reduced sensitivity of magnetic microspheres, dim prospects for industrialization, and complex distillation, precipitation, and polymerization devices.

Active Publication Date: 2021-09-07
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation process of this type of magnetic particles is complicated, and multiple centrifugation operations are required in the preparation process of template microspheres, which is not conducive to industrialization and reduces the cost of use
Polystyrene is a hydrophobic functional material, which will bind non-specifically to proteins in the aqueous phase system, which will greatly reduce the sensitivity and even cause serious consequences of false positives in in vitro diagnostic applications.
[0004] Patent document CN104031201A discloses a preparation method of biological protein magnetic microspheres. The surface modification of the microsphere matrix is ​​realized through emulsion polymerization, thereby obtaining magnetic microspheres coated with a polyacrylate polymer layer. The formation of hydrophilic polymers on the surface of magnetic microspheres reduces the non-specific adsorption of magnetic microspheres to proteins, but this method will reduce the density of functional groups on the surface of functional magnetic microspheres, resulting in a decrease in the sensitivity of magnetic microspheres in in vitro diagnostic applications
[0005] Patent document CN104316679A discloses the application of polyglycidyl ether-modified nano-magnetic microspheres in chemiluminescence immunoassays. The preparation of nano-microspheres with a hydrophilic surface and rich functional groups can inhibit the non-specific adsorption of proteins, but Polyglycidyl ether requires special modification, and the structure is uncontrollable, which is not conducive to large-scale preparation
[0006] Patent document CN104316679A discloses a method for preparing spheres with carboxyl groups. In this method, porous microspheres are prepared first, and then iron salts are precipitated into the porous microspheres to prepare magnetic microspheres. The surface of the magnetic microspheres is modified by distillation, precipitation, and polymerization. , the preparation method in this patent is complicated, the distillation precipitation polymerization device is complicated, and the industrialization prospect is slim

Method used

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  • Surface zwitterionic polymer modified magnetic microsphere as well as preparation method and application of magnetic microsphere
  • Surface zwitterionic polymer modified magnetic microsphere as well as preparation method and application of magnetic microsphere
  • Surface zwitterionic polymer modified magnetic microsphere as well as preparation method and application of magnetic microsphere

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Embodiment 1

[0040] (1) Preparation of magnetic fluid: Weigh 20g of ferrous sulfate and dissolve it in 100ml of ultrapure water, heat up to 70°C, add 10ml of ammonia water, add 5ml of oleic acid after mechanical stirring for 10 minutes, continue stirring for 30min, then heat up to 80°C, After heat preservation for 1 hour, turn off the heating, cool to room temperature, purify, wash with water and methanol, and then dry to obtain oleic acid-modified ferric oxide. The ferroferric oxide modified by oleic acid is uniformly dispersed with chloroform to prepare a magnetic fluid, and the magnetic fluid with a mass concentration of 50% is obtained. figure 1 It is the morphology under the magnetic fluid transmission microscope.

[0041](2) Preparation of submicron magnetic microspheres: accurately weigh 100 mg sodium lauryl sulfate and disperse it in 50 ml ultrapure water, take 2 ml of the magnetic fluid in step (1) and disperse it in the SDS aqueous solution, and use a cell disruptor 200W Sonicat...

Embodiment 2

[0049] (1) Preparation of zwitterion-coated magnetic microspheres: Accurately weigh 100mg of poly(4-styrenesulfonic acid-copolymerization-maleic acid-sodium salt) and add it to 100ml of acetonitrile, add 100mg of the Silicon-coated magnetic microspheres, 100 μl methacrylic acid, 100 mg N,N'-methylenebisacrylamide, 10 mg ethylene glycol methacrylate, 2.5 mg azobisisobutyronitrile, high-purity nitrogen and continue After stirring for 30 minutes, start heating to 90° C., continue to stir and react for 2 hours, then purify, wash with ethanol and disperse into water to obtain zwitterion-modified magnetic microspheres.

[0050] (2) Magnetic microglobulin coupling: Disperse 10 mg of magnetic microspheres in step 4 into 500 μl pH 6.0 MES buffer, mix well, and then magnetically separate once, discard the supernatant, continue to add 400 μl pH 6.0 MES buffer, and add 100 μl EDC Solution, 40ug NT-proBNP antibody, incubated at 37°C for 30 minutes, then magnetically separated, discarded th...

Embodiment 3

[0053] (1) Preparation of magnetic fluid: Weigh 12g of ferrous sulfate heptahydrate and 20g of ferric chloride hexahydrate, dissolve them in 100ml of ultrapure water, heat up to 65°C, add 10ml of ammonia water, stir mechanically for 10 minutes, then add 5ml of oil acid, continue to stir for 30 minutes, then raise the temperature to 80°C, keep warm for 1 hour, turn off the heating, cool to room temperature, purify, wash with water and methanol, and dry to obtain oleic acid-modified ferric oxide. The magnetic fluid was prepared by uniformly dispersing the oleic acid-modified iron ferric oxide with chloroform to obtain a magnetic fluid with a mass concentration of 60%.

[0054] (2) Preparation of submicron magnetic microspheres: Accurately weigh 20 μl tween20 and disperse it into 50 ml ultrapure water, take 2 ml of the magnetic fluid in step (1) and disperse it into the SDS aqueous solution, and use a cell disruptor 200W to sonicate for 10 minutes to obtain the mass Concentration...

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Abstract

The invention relates to a surface zwitterionic polymer modified magnetic microsphere and a preparation method and application thereof, and the preparation method comprises the following steps: (1) preparing magnetic nanoparticles, and dispersing the magnetic nanoparticles in an organic solvent to prepare a magnetic fluid; (2) dispersing the magnetic fluid into a water phase containing a surfactant, and preparing submicron magnetic microspheres by using a reverse microemulsion method; (3) preparing silica modified magnetic nanoparticles; and (4) dispersing in a solvent containing an anionic surfactant, a carboxyl monomer, an amino monomer, a cross-linking agent and an initiator, heating, reacting and polymerizing to obtain the superparamagnetic microspheres coated with the zwitterionic polymer on the surface. The active groups on the surface of the magnetic microsphere can be controlled through the proportion of monomers, biological macromolecules such as nucleic acid and protein can be efficiently and specifically coupled, non-specific binding of the biological macromolecules on the surface of the magnetic microsphere can be inhibited, and the sensitivity and cell sorting efficiency in protein detection are greatly improved.

Description

technical field [0001] The invention relates to the preparation of magnetic microspheres, in particular to a magnetic microsphere modified by a zwitterionic polymer on the surface, a preparation method and application thereof. Background technique [0002] As a solid phase carrier, magnetic microspheres are widely used in the fields of in vitro diagnosis and biochemical separation. Magnetic microspheres are not only used for the capture and manipulation of proteins, nucleic acids, and cells, but can also be used as probes to provide magnetic detection signals. The magnetic microspheres are all carried out in the solution during the capture and manipulation of proteins, nucleic acids, and cells, and only need to use simple magnetic separation to achieve the purpose of use, which greatly optimizes the efficiency and greatly improves the detection sensitivity. Magnetic microspheres have relatively obvious colors, and can transmit signals through colors. Magnetic microspheres a...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/30B01J20/28G01N33/543
CPCB01J20/103B01J20/06B01J20/265B01J20/28009B01J20/28011B01J20/28021G01N33/54326
Inventor 崔大祥刘关高国
Owner SHANGHAI JIAO TONG UNIV
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