Du Zhuo, Yu Yongliang, Wang Jianhua
Research Center for Analytical Sciences, Northeastern University, Box 332, Shenyang, 110004, China.
Anal Bioanal Chem. 2008 Nov;392(5):937-46. doi: 10.1007/s00216-008-2331-2. Epub 2008 Aug 27.
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.
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。