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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Use of quasi-isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances.
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Use of quasi-isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances.

机译：使用准等电缓冲液作为阳极电解液和阴极电解液，以改善毛细管等电聚焦性能。

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摘要

The use of quasi-isoelectric anolytes and catholytes has been investigated to improve CIEF performances. Narrow pH cuts of carrier ampholytes (NC) have been compared to more conventional couples of anolytes/catholytes (phosphoric acid/sodium hydroxide and glutamic acid/lysine). First, a CIEF setup that consists in a bare silica capillary and 70:30 water/glycerol separation medium has been used. The experiments have shown that when using NC instead of more classical anolytes and catholytes, an increase in the protein detection time was observed and the resolutions obtained for neutral and acidic proteins were doubled. Moreover, according to the NC fraction used, the resolution was modified. In order to investigate further the mechanisms involved, a second setup using a capillary coated with hydroxypropylcellulose was used. With this setup no difference has been observed when changing anolyte and catholyte nature. A simple methodology has then been developed to evaluate EOF during focusing and mobilization steps of CIEF experiments. It highlighted the crucial role played by EOF when using a bare silica capillary. EOF indeed decreased by 33% during mobilization step when using NC instead of classical anolytes and catholytes.
机译：已经研究了使用准等电阳极电解液和阴极电解液来改善CIEF性能。已将载体两性电解质（NC）的pH值降低幅度与更常规的阳极电解质/正电解质夫妇（磷酸/氢氧化钠和谷氨酸/赖氨酸）相比。首先，使用了由裸露的硅胶毛细管和70:30的水/甘油分离介质组成的CIEF装置。实验表明，当使用NC代替更经典的阳极电解液和阴极电解液时，观察到蛋白质检测时间增加，并且中性和酸性蛋白质的分离度提高了一倍。此外，根据所用的NC分数，对分辨率进行了修改。为了进一步研究所涉及的机制，使用了第二种装置，该装置使用涂有羟丙基纤维素的毛细管。通过这种设置，在改变阳极电解液和阴极电解液的性质时，没有发现差异。然后开发了一种简单的方法来评估CIEF实验的聚焦和动员步骤中的EOF。它强调了EOF在使用裸硅胶毛细管时所起的关键作用。当使用NC代替经典的阳极电解液和阴极电解液时，在动员步骤中，EOF确实下降了33％。

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