An ultra-stable cytoplasmic antibody engineered for in vivo applications

Hiroyuki Kabayama, Makoto Takeuchi, Naoko Tokushige, Shin ichi Muramatsu, Miyuki Kabayama, Mitsunori Fukuda, Yoshiyuki Yamada, Katsuhiko Mikoshiba

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Targeting cytoplasmic protein–protein interactions with antibodies remains technically challenging, since antibodies expressed in the cytosol frequently form insoluble aggregates. Existing engineering methods are based on the notion that the estimated net charge at pH 7.4 affects stability; as such, they are unable to overcome this problem. Herein, we report a versatile method for engineering an ultra-stable cytoplasmic antibody (STAND), with a strong estimated net negative charge at pH 6.6, by fusing peptide tags with a highly negative charge and a low isoelectric point. Without the need for complicated amino acid substitutions, we convert aggregation-prone antibodies to STANDs that are useful for inhibiting in vivo transmitter release, modulating animal behaviour, and inhibiting in vivo cancer proliferation driven by mutated Kras—long recognised as an “undruggable” oncogenic protein. The STAND method shows promise for targeting endogenous cytoplasmic proteins in basic biology and for developing future disease treatments.

Original languageEnglish
Article number336
JournalNature communications
Volume11
Issue number1
DOIs
Publication statusPublished - 2020 Dec 1

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

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    Kabayama, H., Takeuchi, M., Tokushige, N., Muramatsu, S. I., Kabayama, M., Fukuda, M., Yamada, Y., & Mikoshiba, K. (2020). An ultra-stable cytoplasmic antibody engineered for in vivo applications. Nature communications, 11(1), [336]. https://doi.org/10.1038/s41467-019-13654-9