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Lysine Bioconjugation on Native Albumin with a Sulfonyl Acrylate Reagent

  • Maria J. Matos
  • Gonzalo Jiménez-Osés
  • Gonçalo J. L. BernardesEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2033)

Abstract

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 words

Lysine Sulfonyl acrylate Bioconjugation Aza-Michael addition Human serum albumin 

Notes

Acknowledgments

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Maria J. Matos
    • 1
  • Gonzalo Jiménez-Osés
    • 2
    • 3
  • Gonçalo J. L. Bernardes
    • 1
    • 4
    Email author
  1. 1.Department of ChemistryUniversity of CambridgeCambridgeUK
  2. 2.Departamento de Química, Centro de Investigación en Síntesis QuímicaUniversidad de La RiojaLogroñoSpain
  3. 3.CIC bioGUNEBizkaia Technology ParkDerioSpain
  4. 4.Faculdade de Medicina, Instituto de Medicina MolecularUniversidade de LisboaLisbonPortugal

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