Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

Bernardim, B. ; Cal, P.M.S.D. ; Matos, M.J. ; Oliveira, B.L. ; Martínez-Saéz, N. ; Albuquerque, I.S. ; Perkins, E. ; Corzana, F. ; Burtoloso, A.C.B. ; Jiménez-Osés, G. ; Bernardes, G.J.L.

Resumen: 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.

Idioma: Inglés

DOI: 10.1038/ncomms13128

Año: 2016

Publicado en: Nature Communications 7 (2016), [9 pp.]
ISSN: 2041-1723

Factor impacto JCR: 12.124 (2016)

Categ. JCR: MULTIDISCIPLINARY SCIENCES rank: 3 / 63 = 0.048 (2016) - Q1 - T1

Factor impacto SCIMAGO: 6.413 - Biochemistry, Genetics and Molecular Biology (miscellaneous) (Q1) - Physics and Astronomy (miscellaneous) (Q1) - Chemistry (miscellaneous) (Q1)

Financiación: info:eu-repo/grantAgreement/ES/MICINN/RYC-2013-14706
Financiación: info:eu-repo/grantAgreement/ES/MINECO/CTQ2015-70524-R

Tipo y forma: Artículo (Versión definitiva)