Lysine Bioconjugation on Native Albumin with a Sulfonyl Acrylate Reagent
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This protocol details a novel bioconjugation strategy that uses a methanesulfonyl acrylate reagent that is directed to the most reactive lysine on human serum albumin, which enables the construction of chemically defined and stable bioconjugates. The reaction proceeds rapidly and a regioselective modification is achieved using a single molar equivalent of the reagent under biocompatible conditions (37 °C, pH 8.0). Importantly, the bioconjugate retains both the secondary structural content and function of the unmodified protein. During the reaction of the amino group of lysine and the sulfonyl acrylate reagent, methanesulfinic acid is released after the conjugate addition, which then generates an electrophilic acrylate moiety on the protein. This acrylate can be further used for site-specific protein labeling using a synthetic molecule bearing a reactive amine under biocompatible conditions (21 °C, pH 8.0).
Key wordsLysine Sulfonyl acrylate Bioconjugation Aza-Michael addition Human serum albumin
M.J.M. thanks Xunta da Galicia and Galician Plan of Research, Innovation and Growth 2011–2015 (Plan I2C, ED481B 2014/086-0 and ED481B 2018/007). G.J.O. thanks D.G.I. MINECO/FEDER (CTQ2015-70524-R and RYC-2013-14706 grants). G.J.L.B. is a Royal Society University Research Fellow (URF\R\180019).
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