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Monodisperse core-shell structure polymer nano particle as well as preparation and application thereof

A core-shell structure, nanoparticle technology, applied in material separation, material analysis, other chemical processes, etc., can solve the problems of unfavorable large-scale protein sample pretreatment, low efficiency of post-modification grafting strategy, cumbersome steps, etc. The effect of hydrophilicity, clean and smooth particles, high glycoprotein adsorption capacity

Active Publication Date: 2015-07-01
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some studies have successfully immobilized 3-aminophenylboronic acid on the surface of macroporous silica spheres, ferric oxide particles, and gold nanoparticles (Xu Y, Wu Z, Zhang L, Lu H, Yang P, Webley P.A, Zhao D.Anal .Chem.2009, 81, 503-508; Zhang L, Xu Y, Yao H, Xie L, Yao J, Lu H, Yang P. Chem.--Eur.J.2009, 15, 10158-10166; Tang J , Liu Y, Qi D, Yao G, Deng C, Zhang X.Proteomics.2009, 9, 5046-5055), however, the post-modification grafting strategy adopted is inefficient and the steps are extremely cumbersome
Some studies have prepared boronic acid functionalized monolithic columns by means of copolymerization (L Ren, Y Liu, M Dong, Z. Liu. J. Chromatogr. A. 2009, 1216, 8421-8425; Z Lin, J Pang, H Yang, Z Cai, L Zhang, G. Chen, Chem.Commun.2011, 47, 9675-9677; Y Liu, L Ren, Z Liu.Chem.Commun.2011, 47, 5067-5069), although the boronic acid bonding efficiency is improved And the glycoprotein enrichment effect is better, but the limitation of the column capacity of the monolithic column itself makes this kind of material unfavorable for the pretreatment of large-scale protein samples

Method used

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  • Monodisperse core-shell structure polymer nano particle as well as preparation and application thereof
  • Monodisperse core-shell structure polymer nano particle as well as preparation and application thereof
  • Monodisperse core-shell structure polymer nano particle as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Preparation of monodisperse core-shell polymer nanoparticles

[0029] Such as figure 1 As indicated, weigh 33.8mg of 4-vinylphenylboronic acid (VPBA) and add it into a 50mL round bottom flask containing 2mL of ethanol, and sonicate for 5min. After the monomer was dissolved, 8 mL of water, 212.2 mg of N,N'-methylenebisacrylamide (MBAA) and 5 mg of azobisisobutyronitrile (AIBN) were added to the flask, and the mixture was sonicated until the mixture was completely dissolved. The flask was placed in an oil bath, and the temperature was raised from room temperature to 70° C., and kept stirring and heating at this temperature for 18 hours. After the reaction is finished, the solution is milky, thereby preparing the polymer core.

[0030] Weigh 33.8mg VPBA and 5mg AIBN respectively, and dissolve them in 2mL ethanol by ultrasonic. The solution was added to the polymerization system after the above reaction, stirred and heated at 70° C. for 6 hours, and the solution was s...

Embodiment 2

[0035] 1. Protein mixed solution preparation

[0036] Glycoprotein (horseradish peroxidase, HRP) and non-glycoprotein (bovine serum albumin, BSA) were mixed at a mass ratio of 1:1 and dissolved in 50 mM ammonium bicarbonate buffer solution (pH 9.0) to prepare A protein mixture solution with a concentration of 100 ng / μL was obtained.

[0037] 2. Selective enrichment of glycoproteins

[0038]Weigh 1 mg of core-shell polymer nanoparticles prepared in Example 1, disperse in 200 μL of the above protein mixture solution, shake gently at room temperature for 2 hours, centrifuge and discard the supernatant after the reaction is completed. The material was washed several times with 50 mM ammonium bicarbonate buffer solution (pH 9.0), and the supernatant was discarded by centrifugation. Add 20 μL of 50% acetonitrile solution containing trifluoroacetic acid (1% volume fraction of trifluoroacetic acid) to the separated material, and shake gently at room temperature for 1 hour. The supe...

Embodiment 3

[0043] In order to investigate the selectivity and non-specific adsorption of polymer nanoparticles in the process of glycoprotein enrichment, further interference experiments were carried out. HRP and BSA were mixed at a mass ratio of 1:500 and dissolved in 50 mM ammonium bicarbonate buffer solution (pH 9.0) to prepare a protein mixed solution with a concentration of 25.05 μg / μL. Weigh 1 mg of core-shell polymer nanoparticles, and carry out glycoprotein enrichment and subsequent MALDI-TOF MS identification under the conditions of Example 2.

[0044] Such as Figure 5 As shown, due to the extremely low concentration of HRP in the stock solution, the mass spectrometry response signal was inhibited by BSA and could not be detected (a curve); after enrichment by core-shell polymer nanoparticles, the HRP signal intensity increased significantly (c curve); and excluded The effect of interfering proteins was eliminated, so that BSA was only detected in the supernatant (b curve). I...

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Abstract

The invention belongs to the technical field of high molecular materials and analysis and relates to a monodisperse core-shell structure polymer nano particle as well as preparation and application thereof. The polymer nano particle is a core-shell type copolymer of an acrylic acid cross-linking agent and a monoene phenylboronic acid functional monomer; and the polymer nano particle is prepared by using a one-pot method, that is, precipitation polymerization with the combination of in-situ package strategy. Synthesis steps of the conventional core-shell structure polymer particles are simplified, and the polymer nano particle has the characteristics of rapidness, convenience and easiness, low cost, multiple boric acid functional monomers on the surface, and the like. The polymer nano particle can be used for separating or enriching glycoprotein with a 1,2-cis-glycol structure, and has good practical values and application prospects in the fields of proteomics and the like.

Description

technical field [0001] The invention relates to the enrichment of glycoproteins, in particular to a monodisperse core-shell structure polymer nanoparticle and its preparation and application in glycoprotein enrichment. Background technique [0002] It is well known that the composition of proteins is extremely complex and constantly changing, even in different cells of the same species or in different stages of the same cell, there are significant differences in the composition of proteins. Especially for post-translational modified proteins, they often become hot spots and difficulties in proteomics research because of their low abundance and complexity. [0003] As a class of important post-translational modification proteins, glycoproteins play an important role in biological processes such as cell adhesion, signal transduction, immunity and tumorigenesis. In recent years, although mass spectrometry has become an effective tool for studying glycosylation site information...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F222/38C08F230/06B01J20/281G01N30/06
Inventor 张丽华曲焱焱刘键熙杨开广张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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