A convenient method for preparation of an engineered mouse interleukin-3 analog with high solubility and wild-type bioactivity
- Author(s)
- Murphy, JM; Metcalf, D; Young, IG; Hilton, DJ;
- Details
- Publication Year 2010-04,Volume 28,Issue #2,Page 104-110
- Journal Title
- GROWTH FACTORS
- Publication Type
- Journal Article
- Abstract
- Mouse interleukin-3 (mIL-3) is a critical cytokine regulator of myeloid cell differentiation, survival and activation, and consequently this cytokine has become a key reagent for hematological studies in the laboratory. Although bacterial expression has been used for the preparation of recombinant mIL-3 for more than 20 years, the resultant cytokine is known to exhibit poor solubility, be prone to aggregation, and may contain mispaired disulfide bonds. As a result, little structural characterization of mIL-3 has been possible to date. In the present work, we describe a convenient, inexpensive, and scalable protocol for preparing an mIL-3 analog with wild-type bioactivity from Escherichia coli via a simple purification scheme. This analog is typically expressed at > 1 mg/l of shaking Super broth culture and, owing to solubility > 5 mg/ml, structural studies in solution by nuclear magnetic resonance spectroscopy are feasible for mIL-3 for the first time.
- Publisher
- TAYLOR & FRANCIS LTD
- Keywords
- MURINE INTERLEUKIN-3; BIOLOGIC PROPERTIES; MAST-CELL; PURIFICATION; IL-5; HOMOGENEITY; RECEPTORS; BINDING; GROWTH; FORMS
- Publisher's Version
- https://doi.org/10.3109/08977190903443048
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2010-04-01 12:00:00