Design and Synthesis of a Homogeneous Erythropoietin Analogue with Two Human Complex-Type Sialyloligosaccharides: Combined Use of Chemical and Bacterial Protein Expression Methods

Authors

  • Kiriko Hirano,

    1. International Graduate School of Arts and Sciences, Yokohama City University, 22-2, Seto, Kanazawa-ku, Yokohama, 236-0027 (Japan)
    Search for more papers by this author
  • Derek Macmillan Prof. Dr.,

    1. Department of Chemistry, University College London, 20 Gordon Street, London WC1H 0AJ (UK)
    Search for more papers by this author
  • Katsunari Tezuka Dr.,

    1. Research Project for Glycoprotein Engineering funded by Otsuka Chemical Co., Ltd., Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510 (Japan)
    Search for more papers by this author
  • Takashi Tsuji Prof. Dr.,

    1. Department of Biological Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510 (Japan)
    Search for more papers by this author
  • Yasuhiro Kajihara Prof. Dr.

    1. International Graduate School of Arts and Sciences, Yokohama City University, 22-2, Seto, Kanazawa-ku, Yokohama, 236-0027 (Japan)
    2. Current address: Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043 (Japan)
    Search for more papers by this author

  • Financial support from the Japan Society for the promotion of Science (Grant-in-Aid for Creative Scientific Research no.17GS0420 and 18550155) is acknowledged. We thank Dr. Yukishige Ito (Riken), Dr. Yukio Nishina (Yokohama City University), Yuri Hatakeyama, Kyoko Isumi, Ai Himeno (Otsuka Chemical Co.), and Ayako Kajihara for technical support and encouragement.

Abstract

original image

Highly expressive: Cell proliferation was observed with concentrations of an erythropoietin (EPO) analogue above 50 pg mL−1. The EPO analogue has two human complex-type sialyloligosaccharides (see picture) and was formed by the combined use of chemical synthesis and protein expression in E. coli. Both the 24- and 30-positions are glycosylated, but the two sialyloligosaccharides do not interfere with binding of the EPO analogue to a receptor.

Ancillary