A poly(2-hydroxyethyl methacrylate)-based immobilized metal affinity chromatography adsorbent for protein purification

Journal of The Chinese Institute of Chemical Engineers Pub Date : 2008-07-01 DOI:10.1016/j.jcice.2008.03.005

Che-Liang Liou, Yi-Chuan Chen, Sung-Chyr Lin

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引用次数: 13

Abstract

Poly(HEMA) microbeads were prepared by suspension polymerization of 2-hydorxyethylmethacrylate and ethyleneglycoldimethacrylate (EGDMA). The water content, ligand density, and selectivity for poly(His)-tagged d-hydantoinase of the poly(HEMA)-based adsorbents were affected by the concentration of EDGMA used during polymerization. The Ni(II)-loaded poly(HEMA) adsorbent exhibited an adsorption capacity of 1.0 mg/g for poly(His)-tagged d-hydantoinase under optimal conditions with buffer containing 100–300 mM NaCl at pH 6.0. One-step purification protocol with the adsorbent gave a purity of at least 92%. The adsorption capacity of adsorbent declined by 54% after 7 cycles, due to the leaching of Ni(II) from the adsorbent. However, upon regeneration the adsorption capacity can be restored. Given the ease of preparation and the chemical and microbial resistance, the poly(HEMA)-based IMAC adsorbent could be a promising substitute for the polysaccharide-based IMAC adsorbents.

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一种用于蛋白质纯化的固定化金属亲和层析吸附材料

以2-羟基甲基丙烯酸乙酯和乙二醇二甲基丙烯酸乙酯(EGDMA)为原料，采用悬浮聚合法制备聚HEMA微球。聚(HEMA)基吸附剂的含水量、配体密度和聚(His)标记的d-羟化酶的选择性受聚合过程中EDGMA浓度的影响。负载Ni(II)的poly(HEMA)吸附剂在含100-300 mM NaCl、pH 6.0的缓冲液条件下，对poly(His)标记的d-羟酶的吸附量为1.0 mg/g。吸附剂一步纯化方案纯度至少为92%。经过7次循环后，吸附剂的吸附量下降了54%，主要原因是Ni(II)从吸附剂中浸出。然而，再生后的吸附能力可以恢复。由于制备简单、耐化学和耐微生物，以聚HEMA为基础的IMAC吸附剂有望取代以多糖为基础的IMAC吸附剂。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of The Chinese Institute of Chemical Engineers

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