Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Kyoto-Daigaku Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
Department of Molecular and Cellular Physiology, Graduate School of Medicine, Kyoto University, Sakyo-ku Yoshida Konoe-cho, Kyoto, 606-8501, Japan.
Protein Expr Purif. 2024 Nov;223:106560. doi: 10.1016/j.pep.2024.106560. Epub 2024 Jul 31.
Automation of protein purification methods can increase researchers' efficiency in life sciences. However, currently reported automated protein purification methods require cost-intensive fast protein liquid chromatography systems, such as ÄKTA pure and ÄKTA explorer, without any reported application to the more cost-efficient entry-level system, ÄKTA go. To fill this gap, here we propose a fast, efficient, and versatile automated protein purification strategy for the ÄKTA go. Straightforward integration of two additional accessories, a column valve and a sample loop, into the default ÄKTA go system and making minor rearrangements of flow lines, enabled automation of multi-step protein purification processes. Utilizing this established system, we demonstrate the automated purification of three distinct types of proteins: ubiquitin, polyhistidine-tagged talin, and GST-tagged human rhinovirus 14 3C protease. The described automation strategy is suitable even for small budget-conscious laboratories operating on ÄKTA go systems, thus reducing researchers' time and efforts spent on routine sample preparation tasks of their investigations.
蛋白质纯化方法的自动化可以提高生命科学研究人员的效率。然而，目前报道的自动化蛋白质纯化方法需要成本高昂的快速蛋白质液相色谱系统，如 ÄKTA pure 和 ÄKTA explorer，而没有任何报道将其应用于更具成本效益的入门级系统 ÄKTA go。为了填补这一空白，我们在这里为 ÄKTA go 提出了一种快速、高效和通用的自动化蛋白质纯化策略。只需将两个额外的附件（柱阀和样品环）直接集成到默认的 ÄKTA go 系统中，并对流路进行微小的重新布置，即可实现多步蛋白质纯化过程的自动化。利用这个已建立的系统，我们展示了三种不同类型的蛋白质的自动化纯化：泛素、多组氨酸标记的桩蛋白和 GST 标记的人鼻病毒 14 3C 蛋白酶。所描述的自动化策略甚至适用于预算有限、使用 ÄKTA go 系统的注重成本效益的小型实验室，从而减少研究人员在其研究中常规样品制备任务上花费的时间和精力。
