层层自组装多壁碳纳米管在线固相萃取碱性蛋白质
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摘要
采用层层自组装法将羧基化多壁碳纳米管(c-MWCNTs)固定在石英棉(QW)上,以其作为固相萃取材料,建立了在线固相萃取碱性蛋白质的方法。考察了pH值、洗脱溶剂、洗脱流速和洗脱体积对固相萃取的影响。研究表明,在优化条件下,该吸附剂对血红蛋白(Hb)和溶菌酶(Lyz)的吸附容量分别为36.1μg/mg、28.3μg/mg。当样品溶液体积为3.0mL时,Hb和Lyz的富集倍数分别可达18和28。SDS-聚丙烯酰胺凝胶电泳的结果显示,该萃取体系可实现对血液中Hb和蛋清中Lyz的有效分离富集。
The carboxylated multi-walled carbon nanotubes(c-MWCNTs)coated quartz wool(c-MWCNTs/QW)prepared by layer-by-layer self-assembly was used as solid-phase extraction(SPE)absorbent for on-line separation and preconcentration of basic proteins.A distinct feature of the c-MWCNTs/QW-packed microcolumn provided advantages of minimized flow impedance when operated in a flow system.With a sample loading volume of 3.0 mL,enrichment factors of 18 and 28 were derived for hemoglobin and lysozyme,and the maximum adsorption capacities of hemoglobin and lysozyme were 36.1μg/mg and 28.3μg/mg,respectively.Various parameters affecting SPE efficiency including the volume,pH,ionic strength and flow rate of sample and eluent were systematically studied.The feasibility of the proposed method was validated by successfully separation of lysozyme in egg white and hemoglobin in human whole blood.
The carboxylated multi-walled carbon nanotubes(c-MWCNTs)coated quartz wool(c-MWCNTs/QW)prepared by layer-by-layer self-assembly was used as solid-phase extraction(SPE)absorbent for on-line separation and preconcentration of basic proteins.A distinct feature of the c-MWCNTs/QW-packed microcolumn provided advantages of minimized flow impedance when operated in a flow system.With a sample loading volume of 3.0 mL,enrichment factors of 18 and 28 were derived for hemoglobin and lysozyme,and the maximum adsorption capacities of hemoglobin and lysozyme were 36.1μg/mg and 28.3μg/mg,respectively.Various parameters affecting SPE efficiency including the volume,pH,ionic strength and flow rate of sample and eluent were systematically studied.The feasibility of the proposed method was validated by successfully separation of lysozyme in egg white and hemoglobin in human whole blood.
引文
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