Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

  1. Bernardim, B. 36
  2. Cal, P.M.S.D. 35
  3. Matos, M.J. 3
  4. Oliveira, B.L. 3
  5. Martínez-Saéz, N. 3
  6. Albuquerque, I.S. 5
  7. Perkins, E. 1
  8. Corzana, F. 23
  9. Burtoloso, A.C.B. 6
  10. Jiménez-Osés, G. 24
  11. Bernardes, G.J.L. 35
  1. 1 Albumedix Ltd, Castle Court, 59 Castle Boulevard, Nottingham, United Kingdom
  2. 2 Universidad de La Rioja
    info

    Universidad de La Rioja

    Logroño, España

    ROR https://ror.org/0553yr311

  3. 3 University of Cambridge
    info

    University of Cambridge

    Cambridge, Reino Unido

    ROR https://ror.org/013meh722

  4. 4 Universidad de Zaragoza
    info

    Universidad de Zaragoza

    Zaragoza, España

    ROR https://ror.org/012a91z28

  5. 5 Universidade de Lisboa
    info

    Universidade de Lisboa

    Lisboa, Portugal

    ROR https://ror.org/01c27hj86

  6. 6 Universidade de São Paulo
    info

    Universidade de São Paulo

    São Paulo, Brasil

    ROR https://ror.org/036rp1748

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Revue:
Nature Communications

ISSN: 2041-1723

Année de publication: 2016

Volumen: 7

Type: Article

DOI: 10.1038/NCOMMS13128 SCOPUS: 2-s2.0-84992671846 WoS: WOS:000386218000001 GOOGLE SCHOLAR

D'autres publications dans: Nature Communications

Objectifs de Développement Durable

Résumé

Maleimides remain the reagents of choice for the preparation of therapeutic and imaging protein conjugates despite the known instability of the resulting products that undergo thiol-exchange reactions in vivo. Here we present the rational design of carbonylacrylic reagents for chemoselective cysteine bioconjugation. These reagents undergo rapid thiol Michael-addition under biocompatible conditions in stoichiometric amounts. When using carbonylacrylic reagents equipped with PEG or fluorophore moieties, this method enables access to protein and antibody conjugates precisely modified at pre-determined sites. Importantly, the conjugates formed are resistant to degradation in plasma and are biologically functional, as demonstrated by the selective imaging and detection of apoptotic and HER2+ cells, respectively. The straightforward preparation, stoichiometric use and exquisite cysteine selectivity of the carbonylacrylic reagents combined with the stability of the products and the availability of biologically relevant cysteine-tagged proteins make this method suitable for the routine preparation of chemically defined conjugates for in vivo applications. © 2016 The Author(s).