Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents
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: Article (Published version)

Creative Commons You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.


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