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Method for preparing magnetic polymer microspheres and application thereof

A technology of polymers and microspheres, applied in the field of biochemistry, can solve the problems of affecting enzymatic reactions, fluid manipulation restricts the use of technology, and is difficult to carry out, so as to achieve the effect of high-throughput biochemical reactions

Active Publication Date: 2021-08-27
SHANGHAI TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, when performing many complex biochemical reactions, samples need to be processed in a certain order to initiate, modify or terminate biochemical reactions, but these complex multi-step operations are difficult to perform in droplets
Although a multi-step process can be achieved by manipulating droplets (droplet fusion, droplet reinjection, droplet separation, and sorting), the use of the above methods requires professional knowledge of microfluidic technology, and the complexity of fluid manipulation Sexuality also limits the use of this type of technology
In microbiological experiments, when bacteria or bacterial genetic material are separated and captured in micro-droplets for a series of corresponding biochemical treatments, it is possible to achieve the connection between multi-step experimental operations and to ensure that there is no cross-contamination and external contamination between samples. source pollution, which is quite challenging
For example, when amplifying and analyzing the genetic material of microorganisms, the microorganisms need to be lysed, but the reagents used in this step may affect the subsequent enzymatic reactions, so there are limitations

Method used

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  • Method for preparing magnetic polymer microspheres and application thereof
  • Method for preparing magnetic polymer microspheres and application thereof
  • Method for preparing magnetic polymer microspheres and application thereof

Examples

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Embodiment 1

[0083] A method for preparing magnetic polymer microspheres, the preparation process is as follows figure 2 As shown in a-2c, it includes the following steps:

[0084] 1) Firstly, polyacrylamide soluble gel microspheres were prepared by microfluidic technology, specifically, N,N, HFE-7500 fluorinated oil phase of N',N'-tetramethylethane-1,2-diamine (TEMED) and 2% g / mL surfactant (PEG-PFPE); second flow input channel Input a mixed aqueous solution containing 6% g / mL acrylamide monomer, 0.392% g / mL N,N'-bis(acryloyl) cystamine and 0.6% g / mL ammonium persulfate; the third output of the microfluidic chip Channels collect soluble gel microdroplets. Place the collected droplets at room temperature for 7 hours to form a gel. After forming the gel, de-emulsify the micro-droplets, and then use HFE-7500 fluorinated oil containing 20% ​​v / v 1H, 1H, 2H, 2H-Perfluorooctanol Wash once, wash twice with n-hexane containing 1% v / v Span-80, wash twice with TE buffer containing 0.1% v / v Trit...

Embodiment 2

[0088] A method for preparing magnetic polymer microspheres, the preparation process is as follows figure 2 As shown in a-2c, it includes the following steps:

[0089] 1) Firstly, polyacrylamide soluble gel microspheres were prepared using microfluidic technology, and the preparation process was the same as step 1 of Example 1);

[0090] 2) Using microfluidic technology to prepare double-layer gel microspheres wrapped with a magnetic layer (the outer layer is polyethylene glycol gel wrapped with magnetic nanoparticles, and the inner layer is polyacrylamide gel), specifically, to Such as image 3 The first flow input channel of the microfluidic chip shown contains 0.4% v / v N,N,N',N'-tetramethylethane-1,2-diamine (TEMED) and 2% g / mL surfactant (PEG-PFPE) HFE-7500 fluorinated oil oil phase, the second flow input channel input containing 5% g / mL polyethylene glycol, 0.6% g / mL ammonium persulfate and trioxide The polyethylene glycol gel solution of ferromagnetic nanoparticles;...

Embodiment 3

[0093] A method for preparing magnetic polymer microspheres, the preparation process is as follows figure 2 As shown in a-2c, it includes the following steps:

[0094] 1) First, prepare soluble polyacrylamide gel microspheres using microfluidic technology, and the preparation process is the same as step 1 of Example 1);

[0095] 2) Using microfluidic technology to prepare double-layer gel microspheres wrapped with a magnetic layer (the outer layer is agarose gel wrapped with magnetic nanoparticles, and the inner layer is polyacrylamide gel), specifically, to such as image 3 The first flow input channel of the shown microfluidic chip inputs HFE-7500 fluorinated oil oil phase containing 2% g / mL surfactant (PEG-PFPE); the second flow input channel input contains 2% g / mL Agarose and agarose solution containing ferroferric oxide magnetic nanoparticles (agarose was dissolved completely at 90°C for 30 minutes); the third flow input channel input the soluble gel microspheres prepar...

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Abstract

The invention provides a method for preparing magnetic polymer microspheres and application of the magnetic polymer microspheres. The preparation method comprises the following steps: firstly, preparing a multi-layer polymer micro-droplet of which the outer layer is coated with magnetic nanoparticles by utilizing a droplet generation technology; carrying out curing or semi-curing and demulsification to obtain a multi-layer polymer microsphere coated with a magnetic shell layer; and dissolving at least one layer in the inner-layer polymer of the multi-layer polymer microsphere to obtain the magnetic polymer microsphere containing the hollow cavity. The preparation method provided by the invention can be used in any biochemical reaction, the magnetic polymer microspheres serve as a reaction container, the shell layer of the magnetic polymer microspheres serves as a passive sieve to retain a packaged reaction substrate and isolate invasion of external pollution, and meanwhile, a small-molecule reaction reagent is allowed to diffuse through and enter the microspheres to react with the reaction substrate. According to the invention, high-efficiency and high-flux biochemical reaction can be carried out under the condition of isolating external pollution; and cross contamination and exogenous contamination of samples among multi-step biochemical reactions can be avoided.

Description

technical field [0001] The invention relates to a method for preparing magnetic polymer microspheres and an application thereof, belonging to the technical field of biochemistry. Background technique [0002] With the iterative update of molecular biology techniques, modern molecular biology research increasingly relies on high-throughput analysis methods to process complex samples with single-cell or single-molecule resolution. Compared with microwell plates, capturing single cells, DNA, enzymes or biomolecules in water-in-oil microdroplets or other forms of microscopic compartments can achieve large-scale high-throughput parallel analysis. However, when performing many complex biochemical reactions, samples need to be processed in a certain order to initiate, modify or terminate biochemical reactions, but these complex multi-step operations are difficult to perform in droplets. Although a multi-step process can be achieved by manipulating droplets (droplet fusion, droplet...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J13/02B01J13/14C12N15/10
CPCB01J13/02B01J13/14C12N15/1006Y02A50/30
Inventor 刘一凡李璐瑶张蓉
Owner SHANGHAI TECH UNIV
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