Quaternization of Vinyl/Alkynyl Pyridine Enables Ultrafast Cysteine‐Selective Protein Modification and Charge Modulation. Issue 20 (9th April 2019)

• Record Type: Journal Article
• Authors: Matos, Maria J.; Navo, Claudio D.; Hakala, Tuuli; Ferhati, Xhenti; Guerreiro, Ana; Hartmann, David; Bernardim, Barbara; Saar, Kadi L.; Compañón, Ismael; Corzana, Francisco; Knowles, Tuomas P. J.; Jiménez‐Osés, Gonzalo; Bernardes, Gonçalo J. L.
• Is Part Of: Angewandte Chemie international edition. Volume 58:Issue 20(2019)
• Page Start: 6640
• Page End: 6644
• Publication Date: 2019-04-09
• Subjects: antibody–drug conjugates -- bioconjugation -- cysteine -- microfluidics -- protein modification; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201901405
• Languages: English
• ISSNs: 1433-7851

Abstract

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. Abstract : Quaternize to click : The computation‐inspired discovery of quaternization of the nitrogen of vinyl and alkynyl pyridine reagents to enable selective and ultrafast modification of cysteine at near‐stoichiometric quantities is presented. With this strategy, stable and defined antibody–drug conjugates can be prepared. Furthermore, the electrophoretic mobility of the protein may be modulated through simple introduction of a +1 charge to its overall net charge, as shown using microfluidics.