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提出了一种应用于毛细管筛分电泳中的电动超荷电结合柱头水塞堆积样品的方法,实现了十二烷基磺酸钠-蛋白质复合物的在线富集。一般情况下,电动超荷电方法是一种将电动进样与瞬时等速电泳联用的富集技术。具体过程是,首先在毛细管中注入背景电解质,再注入适量的前导电解质,然后电动进样一段时间。最后注入后导电解质开始瞬时等速电泳及分离的过程。本文在常规的电动超荷电技术基础上,在电动进样之前先注入一段含有聚合物的水塞以进一步提高富集效果。同时,考察了电动超荷电中不同富集方法叠加联用的效果,包括聚合物的筛分效应、结合水塞和不结合水塞的场放大样品进样效果、瞬时等速电泳等。结果表明,由于十二烷基磺酸钠-蛋白质复合物的质荷比接近,电动进样中的进样歧视得到消除,电动超荷电结合含聚合物水塞堆积样品的方法可以无歧视地在线富集十二烷基磺酸钠-蛋白质复合物,检测灵敏度增强1 000倍以上。该方法非常适用于低丰度蛋白质的分析,并可同时提供相对分子质量信息。
A new method, which is used in the capillary electrophoresis of supercapacitors, is to accumulate the samples by electrokinetic supercharger combined with the head water plug. The online enrichment of sodium dodecyl sulfate-protein complexes is achieved. In general, the electric super-charging method is an enrichment technique that combines electric injection with instantaneous constant-rate electrophoresis. The specific process is, first in the capillary injection of background electrolyte, and then injected into the appropriate amount of lead electrolyte, and then electric injection for a period of time. After the final infusion of conductive electrolyte began instantaneous isokinetic electrophoresis and separation process. In this paper, based on the conventional electric super-charging technology, a section of water plug containing polymer is injected before electric injection to further enhance the enrichment effect. In the meantime, the effect of superposition and combination of different enrichment methods in electric superchargers was investigated, including the screening effect of polymer, the sample injection effect of water amplification with water plug and non-water plug, and the transient isokinetic electrophoresis. The results show that injection discrimination in electrokinetic injection is eliminated due to the mass-to-charge ratio of the sodium dodecyl sulfate-protein complex, and that the electrokinetic supercharged method in combination with the sample containing polymer water plug can be used without discrimination On-line enrichment of sodium dodecyl sulfate-protein complexes increases detection sensitivity by more than 1,000-fold. This method is well suited for the analysis of low-abundance proteins and provides both molecular weight information.