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Expression, Purification, and Characterization of a Novel Soluble Form of Human Delta-like-1

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Abstract

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 6× His tag, was expressed in Escherichia coli as inclusion bodies after isopropyl β-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 μg/ml of rhDSL. The rhDSL protein might therefore be a potential supplement for the expansion of UCB-primitive hematopoietic cells.

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Acknowledgment

The work was supported by the Science & Technology Commission of Shanghai Municipality (nos. 075407071 and 06dj14001).

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Authors and Affiliations

  1. Laboratory of Regenerative Medicine, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240, People’s Republic of China

    Mei Zhao, Mingyuan Wu, Lingchen Guo, Junfen Jiang, Weiwei Huang, **aojuan Lin, Zhonghui Zhang, Di **ang, Huili Lu, Shunying Zhu & Wei Han

  2. School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, People’s Republic of China

    Yan Yu

  3. Institute for Transfusion Medicine, Charité-Universitätsmedizin Berlin, Berlin, 13351, Germany

    Anja Moldenhauer

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  1. Mei Zhao
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  2. Mingyuan Wu
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Correspondence to Wei Han.

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Zhao, M., Wu, M., Guo, L. et al. Expression, Purification, and Characterization of a Novel Soluble Form of Human Delta-like-1. Appl Biochem Biotechnol 160, 1415–1427 (2010). https://doi.org/10.1007/s12010-009-8603-2

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