A fast and simple automated multi-step protein purification method for ÄKTA go systems

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2024-07-31 DOI:10.1016/j.pep.2024.106560

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引用次数: 0

Abstract

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.

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用于 ÄKTA go 系统的快速、简单的自动化多步蛋白质纯化方法。

蛋白质纯化方法的自动化可以提高生命科学研究人员的工作效率。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，而没有任何报道称其适用于成本效率更高的入门级系统 ÄKTA go。为了填补这一空白，我们在此提出了一种适用于 ÄKTA go 的快速、高效、多功能自动蛋白质纯化策略。在默认的 ÄKTA go 系统中直接集成了两个额外的附件：一个柱阀和一个样品环，并对流路进行了小幅调整，从而实现了多步蛋白质纯化过程的自动化。利用这套成熟的系统，我们展示了三种不同类型蛋白质的自动化纯化：泛素、多聚组氨酸标记的塔林和 GST 标记的人鼻病毒 14 3C 蛋白酶。所述自动化策略甚至适用于使用 ÄKTA go 系统的预算有限的小型实验室，从而减少了研究人员在常规样品制备任务上花费的时间和精力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.