High yield production of a soluble human interleukin-3 variant from E. coli with wild-type bioactivity and improved radiolabeling properties

dc.contributor.authorHercus, T.
dc.contributor.authorBarry, E.
dc.contributor.authorDottore, M.
dc.contributor.authorMcClure, B.
dc.contributor.authorWebb, A.
dc.contributor.authorLopez, A.
dc.contributor.authorYoung, I.
dc.contributor.authorMurphy, J.
dc.contributor.editorvan Raaij, M.
dc.date.issued2013
dc.description.abstractHuman interleukin-3 (hIL-3) is a polypeptide growth factor that regulates the proliferation, differentiation, survival and function of hematopoietic progenitors and many mature blood cell lineages. Although recombinant hIL-3 is a widely used laboratory reagent in hematology, standard methods for its preparation, including those employed by commercial suppliers, remain arduous owing to a reliance on refolding insoluble protein expressed in E. coli. In addition, wild-type hIL-3 is a poor substrate for radio-iodination, which has been a long-standing hindrance to its use in receptor binding assays. To overcome these problems, we developed a method for expression of hIL-3 in E. coli as a soluble protein, with typical yields of >3mg of purified hIL-3 per litre of shaking microbial culture. Additionally, we introduced a non-native tyrosine residue into our hIL-3 analog, which allowed radio-iodination to high specific activities for receptor binding studies whilst not compromising bioactivity. The method presented herein provides a cost-effective and convenient route to milligram quantities of a hIL-3 analog with wild-type bioactivity that, unlike wild-type hIL‑3, can be efficiently radio-iodinated for receptor binding studies.
dc.description.statementofresponsibilityTimothy R. Hercus, Emma F. Barry, Mara Dottore, Barbara J. McClure, Andrew I. Webb, Angel F. Lopez, Ian G. Young, James M. Murphy
dc.identifier.citationPLoS ONE, 2013; 8(8):e74376-e74376-8
dc.identifier.doi10.1371/journal.pone.0074376
dc.identifier.issn1932-6203
dc.identifier.issn1932-6203
dc.identifier.orcidMcClure, B. [0000-0002-5201-4127]
dc.identifier.orcidLopez, A. [0000-0001-7430-0135]
dc.identifier.urihttp://hdl.handle.net/2440/101294
dc.language.isoen
dc.publisherPublic Library of Science
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1033368
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/565217
dc.relation.granthttp://purl.org/au-research/grants/arc/FT100100100
dc.rights© 2013 Hercus et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
dc.source.urihttps://doi.org/10.1371/journal.pone.0074376
dc.subjectEscherichia coli; interleukin-3; solubility
dc.titleHigh yield production of a soluble human interleukin-3 variant from E. coli with wild-type bioactivity and improved radiolabeling properties
dc.typeJournal article
pubs.publication-statusPublished

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