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Simplification of affinity macroporous monolith microfluidic column synthesis and its ability for protein separation.
Journal of Pharmaceutical and Biomedical Analysis ( IF 3.1 ) Pub Date : 2020-01-08 , DOI: 10.1016/j.jpba.2020.113099
Ashish Khaparde 1 , Kishore K R Tetala 1
Affiliation  

A generic multi-component approach was designed to perform simultaneous in situ polymerization and ligand immobilization to develop affinity porous polymer based chromatography resin in a facile mode. This strategy exploits the regioselective ring opening reaction between epoxy group of monomer and native functional group of ligand (i.e. amine) under aqueous condition (pH 9.7). As a proof-of-concept, reaction of iminodiacetic acid (IDA) with allyl glycidyl ether (AGE) in presence of other monomer (HEMA) and crosslinkers (DATD, PDA) for 4 h via thermal initiation process (temperature of 65 °C) was shown. Successful polymerization (both ex situ &in situ) was confirmed by visual observation, surface morphology of the polymer by scanning electron microscope and ligand immobilization by FT-IR analysis. Chelation of the metal-ion i.e. copper (Cu (II)) with IDA in the monolith showed IgG adsorption capacity (27.8 mg/g monolith) over IDA-monolith without metal-ion. The affinity column has shown efficient capture of high abundant proteins such as IgG, transferrin and albumin from human plasma.

中文翻译:

亲和大孔整体式微流控柱的合成及其蛋白质分离能力的简化。

设计一种通用的多组分方法,以同时进行原位聚合和配体固定,从而以简便的方式开发出基于亲和多孔聚合物的色谱树脂。该策略利用了在水性条件(pH 9.7)下单体的环氧基与配体的天然官能团(即胺)之间的区域选择性开环反应。作为概念验证,通过热引发过程(温度65°C),亚氨基二乙酸(IDA)与烯丙基缩水甘油醚(AGE)在其他单体(HEMA)和交联剂(DATD,PDA)存在下反应4小时)。通过目视观察,通过扫描电子显微镜的聚合物表面形态和通过FT-IR分析的配体固定证实了成功的聚合(非原位和原位)。金属离子的螯合即 与没有金属离子的IDA整体相比,整体中具有IDA的铜(Cu(II))的IgG吸附容量(27.8 mg / g整体)。亲和柱显示了从人血浆中高效捕获高丰度蛋白质(例如IgG,转铁蛋白和白蛋白)的能力。
更新日期:2020-01-09
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