Han Bin, Wang Pingli, Zhang Lihua, Qu Feng, Liang Zhen, Deng Yulin, Zhang Yukui
School of Life Science and Technology, Beijing Institute of Technology, Beijing, China.
Se Pu. 2009 Jul;27(4):383-6.
Microchip free flow electrophoresis (microFFE) is a significant microscale technique for the continuous pre-fractionation and the preparation of valuable biological samples. In our recent work, monolithic polyacylamide (PAM) materials were polymerized in microchamber by ultraviolet (UV) initiated polymerization. With the further immobilization of a stable pH gradient on the monolith, a novel microchip free flow isoelectric focusing (microFF-IEF) with monolithic immobilized pH gradient (M-IPG) materials was developed, by which fluorescein-5-isothiocyanate (FITC) labeled glycin, proline and lysine, with a minimum pI difference of 0.33 units, were well separated with a resolution higher than that performed by traditional microFF-IEF. Our experimental results demonstrate that by microFF-IEF with M-IPG, not only the interference of mobile carrier ampholytes in buffer, usually indispensable in traditional microFF-IEF, on the further separation by other techniques and the identification by mass spectrometry (MS) could be avoided, but also the improved resolution and detection sensitivity could be obtained compared with traditional microFF-IEF. Therefore, such a novel technique might be promising in microscale consecutive separation and preparation of samples.
微芯片自由流电泳（microFFE）是一种用于连续预分离和制备有价值生物样品的重要微尺度技术。在我们最近的工作中，通过紫外（UV）引发聚合在微腔中聚合了整体聚丙烯酰胺（PAM）材料。随着在整体上进一步固定稳定的pH梯度，开发了一种具有整体固定pH梯度（M-IPG）材料的新型微芯片自由流等电聚焦（microFF-IEF）技术，通过该技术，异硫氰酸荧光素（FITC）标记的甘氨酸、脯氨酸和赖氨酸，最小pI差值为0.33个单位，得到了很好的分离，分辨率高于传统的microFF-IEF。我们的实验结果表明，通过具有M-IPG的microFF-IEF，不仅可以避免传统microFF-IEF中通常不可或缺的缓冲液中移动载体两性电解质对其他技术进一步分离和质谱（MS）鉴定的干扰，而且与传统microFF-IEF相比，可以获得更高的分辨率和检测灵敏度。因此，这种新技术在微尺度连续分离和样品制备方面可能具有广阔的前景。
