Polymer microsphere, preparation and applications thereof

A polymer and microsphere technology, applied in the field of glycoprotein enrichment, can solve the problems of cumbersome steps, unfavorable large-scale protein sample pretreatment, and low efficiency of post-modification grafting strategy, achieving high reaction efficiency and selective enrichment Set, polymer particles clean effect

Active Publication Date: 2014-10-01
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the adopted post-modification grafting strategy 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 Improvement and good glycoprotein enrichment effect, 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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  • Polymer microsphere, preparation and applications thereof
  • Polymer microsphere, preparation and applications thereof
  • Polymer microsphere, preparation and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0022] 1. Preparation of monodisperse core-shell polymer nanoparticles

[0023] In a 150 mL round bottom flask, add 80 mL of acetonitrile, add 105 mg of methacrylic acid (MAA), 755 mg of N, N'-methylenebisacrylamide (MBAA) and 17 mg of azobisisobutyronitrile (AIBN), and sonicate for 1 minute , so that the added reagents were dissolved to form a uniform solution, a Dean-Stark receiver was mounted on the flask, and a condenser was connected to it, after which nitrogen gas was passed for 15 minutes, a magnetic stirrer was added, and the magnetic stirrer was maintained at a speed of 300 rad / min. The reaction device was placed in an oil bath and heated evenly and slowly, and the reactor rose from room temperature to 115° C. within 30 minutes. Under the condition of maintaining 115° C., half of the solvent in the reaction system was distilled off within 2 hours, the reaction was stopped, and cooled to room temperature. Then use a high-speed centrifuge to centrifuge at a speed of 10...

Embodiment 2

[0028] 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 (pH9.0), thereby A protein mixture solution with a concentration of 100 ng / μL was prepared.

[0029] Weigh 1 mg of the poly(MBAA-co-MAA)(MBAA-co-APBA) core-shell polymer nanoparticles prepared in Example 1, disperse them in 200 μL of the above protein solution, and incubate at room temperature for 2 hours to react After centrifugation, the supernatant was retained for identification by mass spectrometry. The material was washed several times with 50mM ammonium bicarbonate buffer solution (pH9.0), and the supernatant was discarded by centrifugation. Add 20 μL of acetonitrile:water:trifluoroacetic acid mixed solution with a volume ratio of 50:49:1 to the separated material, incubate at room temperature for 1 hour, and centrifuge to remove the enriched product. The protein stock solution...

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Abstract

The invention relates to a mono-dispersed core-shell structure polymer nano particle, preparation and applications thereof. At first, monoene functional monomers and polyene crosslinking monomers are polymerized to generate polymer particles taken as the cores of the polymer microsphere, and then 3-acrylamino phenylboronic acid (APBA) and polyene crosslinking monomers are covered on the surface of the cores through a reversible addition-fragmentation chain transfer (RAFT) polymerization technology so as to obtain smooth core-shell polymer nano particles containing boronic acid functional groups. Phenylboronic acid functional groups are introduced to the material surface through a RAFT method, thus the defects existing in the conventional post modification method such as tedious steps, low reaction efficiency, and unstable morphology of direct polymerized products are avoided, moreover, the APBA can be more stably polymerized, and the bonding amount of boronic acid on the surface is increased at the same time. The polymer nano particle can be used to separate or enrich glycoprotein in a 1,2-cis-diol structure, and has a very good practical value and application prospect in fields like proteomics, etc.

Description

technical field [0001] The invention relates to enriching glycoproteins, in particular to a monodisperse core-shell structure polymer nanoparticle and its preparation and application in glycoprotein enrichment. Background technique [0002] With the end of human genome sequencing, proteomics has gradually become the focus of attention. As we all know, the composition of proteins is extremely complex and constantly changing in organisms, 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. Among these proteins, post-translational modified proteins often become hot spots and difficulties in proteomics research due to 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 signal transduction, immunity and tumorigenesis. Therefore, the discovery and identifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F267/10C08F222/38C08F220/58C08F220/06B01J20/26B01D15/08G01N1/28G01N1/40C07K1/14C12N9/08
Inventor 张丽华刘键熙杨开广曲焱焱张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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