Get access

Flow-Based Enzymatic Ligation by Sortase A

Authors

  • Rocco L. Policarpo,

    1. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)
    Search for more papers by this author
  • Hansol Kang,

    1. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)
    Search for more papers by this author
  • Dr. Xiaoli Liao,

    1. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)
    Search for more papers by this author
  • Amy E. Rabideau,

    1. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)
    Search for more papers by this author
  • Mark D. Simon,

    1. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)
    Search for more papers by this author
  • Prof. Bradley L. Pentelute

    Corresponding author
    1. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)
    • Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA)

    Search for more papers by this author

  • We are indebted to Alexander Vinogradov, Chi Zhang, Jingjing Ling, Surin Mong, Tatiana Berger, and Richard Chang for technical assistance and helpful discussions throughout the course of this work. We thank R. John Collier for providing some of the laboratory equipment used to perform this study. We are grateful to the MIT UROP program for providing funding to R.L.P. and H.K. and the NSF GRFP for A.E.R. We thank the MIT startup fund, the MIT S.O.S. Reed Fund, the Damon Runyon Cancer Research Foundation Award, the Sontag Foundation Distinguished Scientist Award, and the MIT Deshpande Innovation Grant for B.L.P.

Abstract

Sortase-mediated ligation (sortagging) is a versatile, powerful strategy for protein modification. Because the sortase reaction reaches equilibrium, a large excess of polyglycine nucleophile is often employed to drive the reaction forward and suppress sortase-mediated side reactions. A flow-based sortagging platform employing immobilized sortase A within a microreactor was developed that permits efficient sortagging at low nucleophile concentrations. The platform was tested with several reaction partners and used to generate a protein bioconjugate inaccessible by solution-phase batch sortagging.

Get access to the full text of this article

Ancillary