Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

• Type: Article in International Scientific Journal
• Date: 2016

Authors

Bernardim, B (Author) Other; Cal, PMSD (Author) Other; Matos, MJ (Author) Other; Oliveira, BL (Author) Other; Martinez Saez, N (Author) Other; Albuquerque, IS (Author) Other; Perkins, E (Author) Other; Corzana, F (Author) Other; Burtoloso, ACB (Author) Other; Jimenez Oses, G (Author) Other; Bernardes, GJL (Author) Other

Journal

Title: Nature Communications

Vol. 7

ISSN: 2041-1723

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 62 Citations

Other information

Authenticus ID: P-00M-626

DOI: 10.1038/ncomms13128

Abstract (EN): 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 9