Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, P. R. China.
Electrophoresis. 2011 Oct;32(20):2898-903. doi: 10.1002/elps.201100187. Epub 2011 Sep 16.
In this study, we developed a picoliter-scale partial translational spontaneous injection approach which is suitable for high-speed protein separation under sodium dodecyl sulfate-capillary gel electrophoresis mode. On the basis of this approach, we built a high-speed CE system for protein separation based on a short capillary and slotted-vial array. The system has the advantages of simple structure, ease of building without the requirement of microfabricated devices, convenient operation, and low cost. Under the optimized conditions, picoliter-scale sample plugs (corresponding to ∼65 μm plug length) were obtained, which ensured both the high speed and the high efficiency in protein separation. Five fluorescein isothiocyanate labeled proteins including myoglobin, egg albumin, bovine serum albumin, phosphorylase b, and myosin were separated within 60 s with an effective separation length of 1.5 cm. Theoretical plates per meter ranging from 2.58×10⁵ to 1.28×10⁶ (corresponding to 0.78-3.88 μm plate height) were obtained. The separation speed and separation efficiency of the present system are comparable to those of most microchip-based capillary electrophoresis systems for protein separation. The relative standard deviations of the migration times were in the range of 0.9-1.3% (n=5). Good linear relationships between log relative molecular mass and migration time were obtained in the molecular weigh range of 17,200-500,000, which demonstrate the present system can be applied in protein relative molecular mass determination.
在这项研究中，我们开发了一种皮升尺度的部分平移自发注入方法，该方法适用于十二烷基硫酸钠-胶束电动毛细管电泳模式下的蛋白质高速分离。在此方法的基础上，我们构建了基于短毛细管和开槽管阵列的蛋白质高速 CE 系统。该系统具有结构简单、无需微制造设备即可构建、操作方便、成本低等优点。在优化条件下，获得了皮升尺度的样品塞（对应于约 65μm 的塞长），这确保了蛋白质分离的高速和高效率。在 60s 内分离了 5 种荧光素异硫氰酸酯标记的蛋白质，包括肌红蛋白、卵清蛋白、牛血清白蛋白、磷酸化酶 b 和肌球蛋白，有效分离长度为 1.5cm。理论塔板数每米从 2.58×10⁵到 1.28×10⁶（对应于 0.78-3.88μm 的板高）。该系统的分离速度和分离效率与大多数基于微芯片的毛细管电泳系统相当。迁移时间的相对标准偏差在 0.9-1.3%（n=5）范围内。在 17,200-500,000 的相对分子质量范围内，获得了对数相对分子质量与迁移时间之间的良好线性关系，表明该系统可用于蛋白质相对分子质量的测定。
