Automated multistep column chromatography on AKTA pure system using in-line sample dilution

摘要

Recent advances in instrumentation of fast protein liquid chromatography and the availability of various prepacked 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 AKTA (TM) 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 similar to 58% and a reduction in manual labor time of similar to 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.