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旋转式蛋白纯化仪:一种基于平行制备重力柱工作流程的低成本自动化蛋白质纯化仪。

REVOLVER: A low-cost automated protein purifier based on parallel preparative gravity column workflows.

作者信息

Diep Patrick, Cadavid Jose L, Yakunin Alexander F, McGuigan Alison P, Mahadevan Radhakrishnan

机构信息

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada.

Institute of Biomedical Engineering, University of Toronto, Toronto, Canada.

出版信息

HardwareX. 2022 Mar 11;11:e00291. doi: 10.1016/j.ohx.2022.e00291. eCollection 2022 Apr.

Abstract

Protein purification is a ubiquitous procedure in biochemistry and the life sciences, and represents a key step in the protein production pipeline. The need for scalable and parallel protein purification systems is driven by the demands for increasing the throughput of recombinant protein characterization. Therefore, automating the process to simultaneously handle multiple samples with minimal human intervention is highly desirable, yet there are only a handful of such systems that have been developed, all of which are closed source and expensive. To address this challenge, we present REVOLVER, a 3D-printed programmable protein purification system based on gravity-column workflows and controlled by Arduino boards that can be built for under $130 USD. REVOLVER takes a cell lysate sample and completes a full protein purification process with almost no human intervention and yields results indistinguishable from those obtained by an experienced biochemist when purifying a real-world protein sample. We further present and describe MULTI-VOLVER, a scalable version of the REVOLVER that allows for parallel purification of up to six samples and can be built for under $250 USD. Both systems can help accelerate protein purification and ultimately link them to bio-foundries for protein characterization and engineering.

摘要

蛋白质纯化是生物化学和生命科学中普遍存在的一项操作,是蛋白质生产流程中的关键步骤。对可扩展且并行的蛋白质纯化系统的需求,是由提高重组蛋白表征通量的要求所驱动的。因此,非常希望实现自动化流程,以最少的人工干预同时处理多个样品,但目前仅开发出了少数此类系统,而且这些系统均为闭源且价格昂贵。为应对这一挑战,我们推出了REVOLVER,这是一种基于重力柱工作流程、由Arduino板控制的3D打印可编程蛋白质纯化系统,制造成本低于130美元。REVOLVER取细胞裂解液样本,几乎无需人工干预即可完成完整的蛋白质纯化过程,在纯化实际蛋白质样本时,其产生的结果与经验丰富的生物化学家所获得的结果无异。我们还展示并描述了MULTI-VOLVER,它是REVOLVER的可扩展版本,可并行纯化多达六个样品,制造成本低于250美元。这两种系统都有助于加速蛋白质纯化,并最终将它们与用于蛋白质表征和工程的生物铸造厂相连接。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8bd6/9058827/d37a8e73e717/ga1.jpg

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