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


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Type
Article
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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.

Publication Date
2021-11-09
Online Publication Date
2021-11-09
Acceptance Date
2021-10-18
Keywords
3101 Biochemistry and Cell Biology, 31 Biological Sciences, Biotechnology, Bioengineering, Generic health relevance
Journal Title
Sci Rep
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).