| Expression, purification, and characterization of a novel soluble form of human Delta-like-1. Appl Biochem Biotechnol. 2010 Mar;160(5):1415-27 Authors: Zhao M, Wu M, Guo L, Jiang J, Huang W, Lin X, Zhang Z, Xiang D, Lu H, Zhu S, Yu Y, Moldenhauer A, Han W The notch signaling pathway plays an important role in inhibiting cell differentiation and enhancing the repopulation capability of hematopoietic stem/progenitor ! cells. In this study, we developed rhDSL, a novel soluble form of Notch ligand Delta-like-1, which contains the DSL domain and the N-terminal sequence of the ligand, and investigated its function in ex vivo expansion of human umbilical cord blood (UCB)-primitive hematopoietic cells. The coding sequence for rhDSL was cloned into a pQE30 vector, and the recombinant rhDSL, fused with a 6x His tag, was expressed in Escherichia coli as inclusion bodies after isopropyl beta-D-thiogalactoside induction. After renaturing by dilutions, the protein was purified through anion exchange followed by affinity chromatography. The purity of rhDSL protein was more than 99% with very low endotoxin. In combination with human c-kit ligand, the effect of rhDSL on ex vivo expansion of UCB CD34(+) cells was found to be optimal at 1.5 microg/ml of rhDSL. The rhDSL protein might therefore be a potential supplement for the expansion of UCB-primitive hematopoietic cells. PMID: 19333560 [PubMed ! - indexed for MEDLINE] | 
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