Repository logo
 

An open-source automated PEG precipitation assay to measure the relative solubility of proteins with low material requirement.

Accepted version
Peer-reviewed

Type

Article

Change log

Abstract

The solubility of proteins correlates with a variety of their properties, including function, production yield, pharmacokinetics, and formulation at high concentrations. High solubility is therefore a key requirement for the development of protein-based reagents for applications in life sciences, biotechnology, diagnostics, and therapeutics. Accurate solubility measurements, however, remain challenging and resource intensive, which limits their throughput and hence their applicability at the early stages of development pipelines, when long-lists of candidates are typically available in minute amounts. Here, we present an automated method based on the titration of a crowding agent (polyethylene glycol, PEG) to quantitatively assess relative solubility of proteins using about 200 µg of purified material. Our results demonstrate that this method is accurate and economical in material requirement and costs of reagents, which makes it suitable for high-throughput screening. This approach is freely-shared and based on a low cost, open-source liquid-handling robot. We anticipate that this method will facilitate the assessment of the developability of proteins and make it substantially more accessible.

Description

Keywords

3101 Biochemistry and Cell Biology, 31 Biological Sciences, Biotechnology, Bioengineering, Generic health relevance

Journal Title

Sci Rep

Conference Name

Journal ISSN

2045-2322
2045-2322

Volume Title

11

Publisher

Springer Science and Business Media LLC

Rights

All rights reserved
Sponsorship
Royal Society (URF\R1\201461)
Wellcome Trust (203249/Z/16/Z)
M.O. is a PhD student funded by AstraZeneca. P.S. is a Royal Society University Research Fellow (URF\R1\201461).
Relationships
Is previous version of: