Xu Zhongqi, Ando Tomomi, Nishine Tsutomu, Arai Akihiro, Hirokawa Takeshi
Applied Chemistry, Department of Chemistry and Chemical Engineering, Graduate School of Engineering, Hiroshima University, Higashi-hiroshima, Japan.
Electrophoresis. 2003 Nov;24(21):3821-7. doi: 10.1002/elps.200305625.
A microchip gel electrophoresis (MCGE) method with electrokinetic supercharging (EKS, electrokinetic injection with transient isotachophoresis) on a single channel chip was developed for high-sensitive detection of a standard mixture of six proteins (phosphorylase b, albumin, ovalbumin, carbonic anhydrase, trypsin inhibitor, and alpha-lactalbumin) in the form of sodium dodecyl sulfate (SDS) complexes. An average lower limit of detectable concentration (LLDC) achieved using UV detection at 214 nm was 0.27 microg/mL that is 30 times lower than that of conventional MCGE on a cross geometry chip. The calibration curves for molecular weight and concentration of SDS-protein complexes suggested that the present EKS-MCGE method had a better linear dynamic range and benefited future applications for qualitative and quantitative analysis of unknown protein samples. It was found that an excessive amount of unbound SDS in the sample deteriorated the preconcentration effect and resolution. The developed method appears greatly promising for high-speed and high sensitive analysis of SDS-proteins by MCGE.
开发了一种在单通道芯片上采用电动增压（EKS，即具有瞬态等速电泳的电动进样）的微芯片凝胶电泳（MCGE）方法，用于高灵敏度检测六种蛋白质（磷酸化酶b、白蛋白、卵清蛋白、碳酸酐酶、胰蛋白酶抑制剂和α-乳白蛋白）的十二烷基硫酸钠（SDS）复合物标准混合物。使用214nm紫外检测实现的平均可检测浓度下限（LLDC）为0.27μg/mL，这比交叉几何形状芯片上的传统MCGE低30倍。SDS-蛋白质复合物分子量和浓度的校准曲线表明，目前的EKS-MCGE方法具有更好的线性动态范围，有利于未来对未知蛋白质样品进行定性和定量分析。发现样品中过量的未结合SDS会降低预浓缩效果和分辨率。所开发的方法对于通过MCGE对SDS-蛋白质进行高速和高灵敏度分析似乎非常有前景。
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