Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/101294
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Full metadata record
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dc.contributor.author | Hercus, T. | - |
dc.contributor.author | Barry, E. | - |
dc.contributor.author | Dottore, M. | - |
dc.contributor.author | McClure, B. | - |
dc.contributor.author | Webb, A. | - |
dc.contributor.author | Lopez, A. | - |
dc.contributor.author | Young, I. | - |
dc.contributor.author | Murphy, J. | - |
dc.contributor.editor | van Raaij, M. | - |
dc.date.issued | 2013 | - |
dc.identifier.citation | PLoS One, 2013; 8(8):e74376-e74376-8 | - |
dc.identifier.issn | 1932-6203 | - |
dc.identifier.issn | 1932-6203 | - |
dc.identifier.uri | http://hdl.handle.net/2440/101294 | - |
dc.description.abstract | Human 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.statementofresponsibility | Timothy R. Hercus, Emma F. Barry, Mara Dottore, Barbara J. McClure, Andrew I. Webb, Angel F. Lopez, Ian G. Young, James M. Murphy | - |
dc.language.iso | en | - |
dc.publisher | Public Library of Science | - |
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.uri | http://dx.doi.org/10.1371/journal.pone.0074376 | - |
dc.subject | Escherichia coli; interleukin-3; solubility | - |
dc.title | High yield production of a soluble human interleukin-3 variant from E. coli with wild-type bioactivity and improved radiolabeling properties | - |
dc.type | Journal article | - |
dc.identifier.doi | 10.1371/journal.pone.0074376 | - |
dc.relation.grant | http://purl.org/au-research/grants/nhmrc/1033368 | - |
dc.relation.grant | http://purl.org/au-research/grants/nhmrc/565217 | - |
dc.relation.grant | http://purl.org/au-research/grants/arc/FT100100100 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | McClure, B. [0000-0002-5201-4127] | - |
dc.identifier.orcid | Lopez, A. [0000-0001-7430-0135] | - |
Appears in Collections: | Aurora harvest 7 Medicine publications |
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hdl_101294.pdf | Published version | 97.15 kB | Adobe PDF | View/Open |
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