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离子液体-聚氯乙烯离聚物用于高选择性分离碱性蛋白质。

Ionic liquid-polyvinyl chloride ionomer for highly selective isolation of basic proteins.

机构信息

Research Center for Analytical Sciences, Box 332, Chemistry Building, Northeastern University, Shenyang 110004, China.

出版信息

Talanta. 2010 Apr 15;81(1-2):637-42. doi: 10.1016/j.talanta.2009.12.059. Epub 2010 Jan 11.

DOI:10.1016/j.talanta.2009.12.059
PMID:20188975
Abstract

Hydrophilic ionic liquid-polyvinyl chloride (PVC) hybrids were prepared by immobilizing N-methylimidazole (N-mim) to PVC chains in toluene. The NmimCl-PVC hybrids were characterized by FT-IR, (1)H NMR, surface charge analysis and elemental analysis. The immobilization ratio, i.e., the percentage of chloride on PVC chain reacting with N-mim to form the hybrid, varies from 4.3% to 15.1% by increasing the N-mim/PVC molar ratio. The most distinct feature of the hybrid is its excellent selectivity for adsorbing basic proteins by effective suppression of the non-specific protein adsorption by pure PVC, and a higher immobilization ratio facilitates better selectivity. In Tris-HCl buffer, 100 microg mL(-1) of basic proteins, i.e., lysozyme (Lys), cytochrome c (cyt-c) and hemoglobin (Hb), were favorably adsorbed with efficiencies of 97%, 98% and 94% by the hybrid with an immobilization ratio of 15.1%, while the adsorption of acidic proteins, i.e., bovine albumin serum (BSA), transferring (Trf) and immunoglobulin G (IgG) were negligible. The retained Lys, cyt-c and Hb could be readily recovered by elution with phosphate buffer, carbonate buffer and SDS solution with efficiencies of 89%, 87% and 84%, respectively. Another feature of the hybrid is the significant improvement of the biocompatibility characterized by the maintenance of the activity of hemoglobin after adsorption and elution process. The practical usefulness of the hybrid was demonstrated by selective isolation of hemoglobin from human whole blood.

摘要

亲水性离子液体-聚氯乙烯 (PVC) 杂化材料是通过将 N-甲基咪唑 (N-mim) 固定在甲苯中的 PVC 链上制备的。NmimCl-PVC 杂化物通过傅里叶变换红外光谱 (FT-IR)、(1)H NMR、表面电荷分析和元素分析进行了表征。通过增加 N-mim/PVC 摩尔比,固定化比,即 PVC 链上与 N-mim 反应形成杂化物的氯的百分比,从 4.3%变化到 15.1%。该杂化物的最显著特征是通过有效抑制纯 PVC 对非特异性蛋白质吸附,对吸附碱性蛋白质具有优异的选择性,且较高的固定化比有利于更好的选择性。在 Tris-HCl 缓冲液中,固定化比为 15.1%的杂化物对 100μg mL(-1)的碱性蛋白质(即溶菌酶 (Lys)、细胞色素 c (cyt-c) 和血红蛋白 (Hb))表现出良好的吸附效率,分别为 97%、98%和 94%,而对酸性蛋白质(即牛血清白蛋白 (BSA)、转铁蛋白 (Trf) 和免疫球蛋白 G (IgG))的吸附可忽略不计。保留的 Lys、cyt-c 和 Hb 可以通过磷酸盐缓冲液、碳酸盐缓冲液和 SDS 溶液洗脱,洗脱效率分别为 89%、87%和 84%。该杂化物的另一个特点是生物相容性得到显著改善,表现在吸附和洗脱过程后血红蛋白活性得以保持。该杂化物的实际用途通过从人全血中选择性分离血红蛋白得到了证明。

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