High Yield Production of a Soluble Human Interleukin-3 Variant from E. coli with Wild-Type Bioactivity and Improved Radiolabeling Properties
Timothy R Hercus, Emma F Barry, Mara Dottore, Barbara J McClure, Andrew I Webb, Angel F Lopez, Ian G Young, James M Murphy
PLOS ONE | PUBLIC LIBRARY SCIENCE | Published : 2013
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..View full abstract
Awarded by National Health and Medical Research Council of Australia (NHMRC)
Awarded by Australian Research Council fellowship
Awarded by NHMRC IRIISS grant
This work was supported by National Health and Medical Research Council of Australia (NHMRC) grants (1033368 and 565217); an Australian Research Council fellowship (FT100100100) to JMM; with additional support from the Victorian State Government Operational Infrastructure Support and NHMRC IRIISS grant (361646). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.