Riek Uwe, Ramirez Sacnicte, Wallimann Theo, Schlattner Uwe
INSERM U884, Université Joseph Fourier, Laboratory of Fundamental and Applied Bioenergetics, Grenoble, France.
Biotechniques. 2009 May;46(6):ix-xii. doi: 10.2144/000113130.
The standard Akta Explorer high-performance liquid chromatography (HPLC) system has limitations for the automation of multidimensional protein purification. Here, we describe simple modifications that allow for automated multidimensional purification protocols to extend the possibilities of the Akta three-dimensional purification kit in terms of column number, flexibility of volumes stocked for re-injection of samples, and available choice of buffers. These modifications do not preclude the use of standard one-dimensional purification protocols. Additionally, we demonstrate a technology for encrypted full remote control of the machine over the Internet by cost-effective use of standard asymmetric digital subscriber line (ADSL) that enables direct remote interaction with the machine without preventing local control. A 4-column purification scheme, including equilibration and cleaning in place (CIP) procedures, was implemented on such a system. It significantly increased reproducibility and shortened processing time by 85%, as compared with manual operation, thus allowing for automated protein purification overnight.
标准的艾克塔探索者高效液相色谱（HPLC）系统在多维蛋白质纯化自动化方面存在局限性。在此，我们描述了一些简单的改进方法，这些方法可实现自动化多维纯化方案，从而在柱数、样品再注入的储备体积灵活性以及可用缓冲液选择方面扩展艾克塔三维纯化试剂盒的可能性。这些改进并不妨碍使用标准的一维纯化方案。此外，我们展示了一种通过经济高效地使用标准非对称数字用户线路（ADSL）在互联网上对机器进行加密全远程控制的技术，该技术能够与机器进行直接远程交互，同时不影响本地控制。在这样的系统上实施了包括平衡和在位清洗（CIP）程序的四柱纯化方案。与手动操作相比，该方案显著提高了重现性，并将处理时间缩短了85%，从而实现了蛋白质的过夜自动化纯化。
