An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter

Kineavy, Florence B, Davies, Alex A, Mitchell, Molly R, Lay, Daniel, Dominguez, Matt J and Stollar, Elliott J ORCID: 0000-0001-9895-0335 (2021) An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2021 (171).

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Abstract

Protein purification is imperative to the study of protein structure and function and is usually used in combination with biophysical techniques. It is also a key component in the development of new therapeutics. The evolving era of functional proteomics is fueling the demand for high-throughput protein purification and improved techniques to facilitate this. It was hypothesized that a multi column plate adaptor (MCPA) can interface multiple chromatography columns of different resins with multi-well plates for parallel purification. This method offers an economical and versatile method of protein purification that can be used under gravity or vacuum, rivaling the speed of an automated system. The MCPA can be used to recover milligram yields of protein by an affordable and time efficient method for subsequent characterization and analysis. The MCPA has been used for high-throughput affinity purification of SH3 domains. Ion exchange has also been demonstrated via the MCPA to purify protein post Ni-NTA affinity chromatography, indicating how this system can be adapted to other purification types. Due to its setup with multiple columns, individual customization of parameters can be made in the same purification, unachievable by the current plate-based methods.

Item Type:	Article
Uncontrolled Keywords:	Proteins, Chromatography, Affinity, Proteomics, Vacuum
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology > School of Life Sciences
Depositing User:	Symplectic Admin
Date Deposited:	21 Jun 2021 10:42
Last Modified:	21 May 2023 01:30
DOI:	10.3791/62075
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3126844