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Quaternization of Vinyl/Alkynyl Pyridine Enables Ultrafast Cysteine-Selective Protein Modification and Charge Modulation.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Matos, Maria J 
Navo, Claudio D 
Hakala, Tuuli 
Ferhati, Xhenti 
Guerreiro, Ana 

Abstract

Quaternized vinyl- and alkynyl-pyridine reagents were shown to react in an ultrafast and selective manner with several cysteine-tagged proteins at near-stoichiometric quantities. We have demonstrated that this method can effectively create a homogenous antibody-drug conjugate that features a precise drug-to-antibody ratio of 2, which was stable in human plasma and retained its specificity towards Her2+ cells. Finally, the developed warhead introduces a +1 charge to the overall net charge of the protein, which enabled us to show that the electrophoretic mobility of the protein may be tuned through the simple attachment of a quaternized vinyl pyridinium reagent at the cysteine residues. We anticipate the generalized use of quaternized vinyl- and alkynyl-pyridine reagents not only for bioconjugation, but also as warheads for covalent inhibition and as tools to profile cysteine reactivity.

Description

Keywords

antibody-drug conjugates, bioconjugation, cysteine, microfluidics, protein modification

Journal Title

Angew Chem Int Ed Engl

Conference Name

Journal ISSN

1433-7851
1521-3773

Volume Title

58

Publisher

Wiley

Rights

All rights reserved
Sponsorship
European Research Council (676832)
Royal Society (URF\R\180019)
European Commission Horizon 2020 (H2020) Marie Sk?odowska-Curie actions (675007)