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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Online concentration and separation of basic proteins using a cationic polyelectrolyte in the presence of reversed electroosmotic flow.
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Online concentration and separation of basic proteins using a cationic polyelectrolyte in the presence of reversed electroosmotic flow.

机译：在反向电渗流的存在下，使用阳离子聚电解质在线浓缩和分离碱性蛋白。

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摘要

We report an online concentration and separation method for basic proteins using poly(diallyldimethylammonium chloride) (PDDA) solutions in the presence of reversed EOF. Using a capillary dynamically coated with 2% PDDA containing 0.1 M NaCl and filled with 1.2% PDDA under neutral conditions (10 mM phosphate, pH 7.0), we have demonstrated the separation of six basic proteins with peak efficiencies ranging from 175 000 to 616 000 plates/m and RSDs of migration time less than 0.4%. Additionally, high-speed separation of six basic proteins (<7 min) was achieved using a short capillary filled with 0.6% PDDA solutions. Under injection of the large-volume sample (210 nL), the LODs at S/N of 3 for basic proteins are down to nanomolar range. For example, the LOD for lysozyme is 1.2 nM, which is a 260-fold sensitivity enhancement compared with conventional injection method. The proposed method has been applied to the stacking of lysozyme in human saliva samples. Without any pretreatment, we also demonstrated the capability of this method to detect low amounts of peptide samples through the stacking of tryptic peptide of myoglobin. The experimental results indicate that our proposed method has great potential for use in clinical diagnosis and proteomics applications.
机译：我们报告了在反向EOF存在下使用聚二烯丙基二甲基氯化铵（PDDA）溶液对碱性蛋白质进行在线浓缩和分离的方法。使用动态涂有2％PDDA的毛细管，其中PDDA含0.1 M NaCl，在中性条件下（10 mM磷酸盐，pH 7.0）填充1.2％PDDA，我们证明了六种碱性蛋白的分离，其峰值效率为175 000至616 000板/米和迁移时间的RSD小于0.4％。此外，使用装有0.6％PDDA溶液的短毛细管可以快速分离6种碱性蛋白质（<7分钟）。在进样大体积样品（210 nL）的情况下，碱性蛋白的S / N为3的LOD降至纳摩尔范围。例如，溶菌酶的LOD为1.2 nM，与常规注射方法相比，灵敏度提高了260倍。拟议的方法已应用于人类唾液样品中溶菌酶的堆叠。在没有任何预处理的情况下，我们还证明了该方法通过堆积肌红蛋白的胰蛋白酶消化肽来检测少量肽样品的能力。实验结果表明，我们提出的方法在临床诊断和蛋白质组学应用中具有巨大的潜力。

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