Tetrazine ligation: fast bioconjugation based on inverse-electron-demand Diels-Alder reactivity

J Am Chem Soc. 2008 Oct 15;130(41):13518-9. doi: 10.1021/ja8053805. Epub 2008 Sep 18.


Described is a bioorthogonal reaction that proceeds with unusually fast reaction rates without need for catalysis: the cycloaddition of s-tetrazine and trans-cyclooctene derivatives. The reactions tolerate a broad range of functionality and proceed in high yield in organic solvents, water, cell media, or cell lysate. The rate of the ligation between trans-cyclooctene and 3,6-di-(2-pyridyl)-s-tetrazine is very rapid (k2 2000 M-1 s-1). This fast reactivity enables protein modification at low concentration.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromatography, High Pressure Liquid
  • Cyclooctanes / chemical synthesis
  • Cyclooctanes / chemistry
  • Electrons*
  • Molecular Structure
  • Spectrometry, Mass, Electrospray Ionization
  • Tetrazoles / chemical synthesis*
  • Tetrazoles / chemistry
  • Time Factors


  • Cyclooctanes
  • Tetrazoles
  • cyclooctane