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用聚二烯丙基二甲基氯化铵功能化的多壁碳纳米管薄层对酸性蛋白质进行选择性分离。

Selective isolation of acidic proteins with a thin layer of multiwalled carbon nanotubes functionalized with polydiallyldimethylammonium chloride.

作者信息

Du Zhuo, Yu Yongliang, Wang Jianhua

机构信息

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

出版信息

Anal Bioanal Chem. 2008 Nov;392(5):937-46. doi: 10.1007/s00216-008-2331-2. Epub 2008 Aug 27.

Abstract

The selective isolation of acidic proteins using a thin layer of multiwalled carbon nanotubes (MWNTs) functionalized with polydiallyldimethylammonium chloride (PDDA) was demonstrated. A certain amount (20 ml) of a suspension of PDDA-functionalized MWNTs that had been well dispersed by sonication was filtered through an MF-Millipore membrane with a pore aperture of 1.2 microm, and a uniform layer of PDDA-MWNT composites with a thickness of ca. 5 microm formed on the membrane. A 4 x 1 cm piece of the obtained membrane was supported by a stainless steel wire mesh and was then sandwiched between two PTFE films with grooved flow-through channels to form an extraction module. This module with a flow inlet and outlet was incorporated into a sequential injection system for performing the on-line separation and preconcentration of acidic protein, i.e., bovine serum albumin (BSA), and the BSA retained on the layer was eluted with a citrate buffer used as stripping reagent. In addition to a significant reduction in flow resistance, a dynamic sorption capacity of 3.8 mg mg(-1) or 1.4 mg cm(-2) for BSA was achieved using the layer-based system--a 146-fold improvement over that obtained using a packed microcolumn mode. A sample volume of 2.0 ml yielded an enrichment factor of 17, a retention efficiency of 100% and a recovery of 95%, along with a sampling frequency of 20 h(-1) and a RSD value of 2.8% at 25 microg ml(-1) for BSA. The practical applicability of the system was demonstrated by isolating acidic proteins (especially human serum albumin) from whole blood.

摘要

已证明使用用聚二烯丙基二甲基氯化铵(PDDA)功能化的多壁碳纳米管(MWNTs)薄层可选择性分离酸性蛋白质。将一定量(20毫升)经超声充分分散的PDDA功能化MWNTs悬浮液通过孔径为1.2微米的MF-密理博膜过滤,在膜上形成一层厚度约为5微米的均匀PDDA-MWNT复合材料层。将得到的膜切成4×1厘米的片,用不锈钢丝网支撑,然后夹在两个带有凹槽流通通道的聚四氟乙烯薄膜之间,形成一个萃取模块。这个带有流体入口和出口的模块被整合到一个顺序注射系统中,用于在线分离和预浓缩酸性蛋白质,即牛血清白蛋白(BSA),保留在该层上的BSA用用作洗脱剂的柠檬酸盐缓冲液洗脱。除了显著降低流动阻力外,使用基于层的系统对BSA的动态吸附容量达到3.8毫克/毫克(-1)或1.4毫克/平方厘米,比使用填充微柱模式提高了146倍。2.0毫升的样品体积产生了17的富集因子、100%的保留效率和95%的回收率,以及25微克/毫升的BSA在20小时(-1)的采样频率和2.8%的相对标准偏差值。通过从全血中分离酸性蛋白质(特别是人血清白蛋白)证明了该系统的实际适用性。

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