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

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cam.issuedOnline2021-11-09
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cam.orpheus.successTue Feb 01 19:02:04 GMT 2022 - Embargo updated
dc.contributor.authorOeller, Marc
dc.contributor.authorSormanni, Pietro
dc.contributor.authorVendruscolo, Michele
dc.contributor.orcidOeller, Marc [0000-0003-0597-5950]
dc.contributor.orcidSormanni, Pietro [0000-0002-6228-2221]
dc.contributor.orcidVendruscolo, Michele [0000-0002-3616-1610]
dc.date.accessioned2021-10-19T23:31:36Z
dc.date.available2021-10-19T23:31:36Z
dc.date.issued2021-11-09
dc.description.abstractThe 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.
dc.description.sponsorshipM.O. is a PhD student funded by AstraZeneca. P.S. is a Royal Society University Research Fellow (URF\R1\201461).
dc.identifier.doi10.17863/CAM.77096
dc.identifier.eissn2045-2322
dc.identifier.issn2045-2322
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/329647
dc.languageeng
dc.language.isoeng
dc.publisherSpringer Science and Business Media LLC
dc.publisher.urlhttp://dx.doi.org/10.1038/s41598-021-01126-4
dc.rightsAll rights reserved
dc.subject3101 Biochemistry and Cell Biology
dc.subject31 Biological Sciences
dc.subjectBiotechnology
dc.subjectBioengineering
dc.subjectGeneric health relevance
dc.titleAn open-source automated PEG precipitation assay to measure the relative solubility of proteins with low material requirement.
dc.typeArticle
dcterms.dateAccepted2021-10-18
prism.issueIdentifier1
prism.publicationDate2021
prism.publicationNameSci Rep
prism.startingPage21932
prism.volume11
pubs.funder-project-idRoyal Society (URF\R1\201461)
pubs.funder-project-idWellcome Trust (203249/Z/16/Z)
rioxxterms.licenseref.startdate2021-11-09
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review
rioxxterms.versionAM
rioxxterms.versionofrecord10.1038/s41598-021-01126-4

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