Abstract
Recent advances in instrumentation of fast protein liquid chromatography and the availability of various pre-packed columns helped to standardize protein purification protocols. However, purification processes are often plagued by inefficient manual procedures between the column purification steps, such as sample transfer, dialysis, concentration and filtration. Working with automation development in lab scale aims at giving immediate increase in research productivity. Here, we describe the automation of a four-column step purification using a modular sample dilution process coupled with ion exchange chromatography on a commonly available ÄKTA™ pure chromatography system. The method was applied to purify detergent-solubilized G protein-coupled receptors. Queuing multiple samples resulted in reduction in experiment time of up to ~58% and a reduction in manual labor time of ~83% while sustaining a comparable protein yield. The resultant combination of in-line dilution and ion-exchange chromatography could function as a generic method for automating other multistep column purifications schemes.
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