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A versatile multidimensional protein purification system with full Internet remote control based on a standard HPLC system

2009; Future Science Ltd; Volume: 46; Issue: sup6 Linguagem: Inglês

10.2144/000113130

1940-9818

Uwe Riek, Sacnicte Ramirez, Theo Wallimann, Uwe Schlattner,

Glycosylation and Glycoproteins Research

The standard Äkta 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 Äkta 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.