000057762 001__ 57762
000057762 005__ 20200221144339.0
000057762 0247_ $$2doi$$a10.1038/ncomms13128
000057762 0248_ $$2sideral$$a96993
000057762 037__ $$aART-2016-96993
000057762 041__ $$aeng
000057762 100__ $$aBernardim, B.
000057762 245__ $$aStoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents
000057762 260__ $$c2016
000057762 5060_ $$aAccess copy available to the general public$$fUnrestricted
000057762 5203_ $$aMaleimides 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.
000057762 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/CTQ2015-70524-R$$9info:eu-repo/grantAgreement/ES/MICINN/RYC-2013-14706
000057762 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000057762 590__ $$a12.124$$b2016
000057762 591__ $$aMULTIDISCIPLINARY SCIENCES$$b3 / 63 = 0.048$$c2016$$dQ1$$eT1
000057762 592__ $$a6.413$$b2016
000057762 593__ $$aBiochemistry, Genetics and Molecular Biology (miscellaneous)$$c2016$$dQ1
000057762 593__ $$aPhysics and Astronomy (miscellaneous)$$c2016$$dQ1
000057762 593__ $$aChemistry (miscellaneous)$$c2016$$dQ1
000057762 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000057762 700__ $$aCal, P.M.S.D.
000057762 700__ $$aMatos, M.J.
000057762 700__ $$aOliveira, B.L.
000057762 700__ $$aMartínez-Saéz, N.
000057762 700__ $$aAlbuquerque, I.S.
000057762 700__ $$aPerkins, E.
000057762 700__ $$aCorzana, F.
000057762 700__ $$aBurtoloso, A.C.B.
000057762 700__ $$0(orcid)0000-0003-0105-4337$$aJiménez-Osés, G.
000057762 700__ $$aBernardes, G.J.L.
000057762 773__ $$g7 (2016), [9 pp.]$$pNATURE COMMUNICATIONS$$tNature Communications$$x2041-1723
000057762 8564_ $$s3317653$$uhttps://zaguan.unizar.es/record/57762/files/texto_completo.pdf$$yVersión publicada
000057762 8564_ $$s68039$$uhttps://zaguan.unizar.es/record/57762/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000057762 909CO $$ooai:zaguan.unizar.es:57762$$particulos$$pdriver
000057762 951__ $$a2020-02-21-13:48:56
000057762 980__ $$aARTICLE