Measurement of Sialic Acid Content Is Insufficient to Assess Bioactivity of Recombinant Human Erythropoietin

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  • Yanagihara Shigehiro
    Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd. Graduate School of Natural Science and Technology, Okayama University
  • Taniguchi Yuya
    Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd.
  • Hosono Mareto
    Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd.
  • Yoshioka Eiji
    Pharmacokinetic Research Laboratories, Research Division, Kyowa Hakko Kirin Co., Ltd.
  • Ishikawa Rika
    Takasaki Plant, Production Division, Kyowa Hakko Kirin Co., Ltd.
  • Shimada Yoshihiro
    Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd.
  • Kadoya Toshihiko
    Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd.
  • Kutsukake Kazuhiro
    Graduate School of Natural Science and Technology, Okayama University

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Assessment of biological potency and its comparison with clinical effects are important in the quality control of therapeutic glycoproteins. Animal models are usually used for evaluating bioactivity of these compounds. However, alternative methods are required to simplify the bioassay and avoid ethical issues associated with animal studies. Negatively charged sialic acid residues are known to be critical for in vivo bioactivity of recombinant human erythropoietin (rhEPO). In this study, we used capillary zone electrophoresis, a charge-based separation method, to estimate the sialic acid content for predicting in vivo bioactivity of rhEPO. In vivo bioactivities of rhEPO subfractions were measured and compared with sialylation levels. The results obtained indicated that in vivo bioactivity of rhEPO is not simply correlated with the sialylation level, which suggests that it is difficult to predict biological potency from the sialic acid content alone. N-Glycan moieties as well as sialic acid residues may have a significant impact on in vivo bioactivity of rhEPO.

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