Template-directed covalent conjugation of DNA to native antibodies, transferrin and other metal-binding proteins

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  • Christian B Rosen, 1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Chemistry, Langelandsgade 140, 8000 Aarhus C, Denmark [3]., Danmark
  • Anne L B Kodal, 1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Chemistry, Langelandsgade 140, 8000 Aarhus C, Denmark [3]., Danmark
  • Jesper S Nielsen
  • David H Schaffert, 1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Molecular Biology and Genetics, Aarhus University, C. F. Møllers Allé 3, 8000 Aarhus C, Denmark., Danmark
  • Carsten Scavenius
  • Anders H Okholm, 1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Molecular Biology and Genetics, Aarhus University, C. F. Møllers Allé 3, 8000 Aarhus C, Denmark., Danmark
  • Niels V Voigt, 1] Center for DNA Nanotechnology and Interdisciplinary Nanoscience Center, Gustav Wieds Vej 14, 8000 Aarhus C, Denmark [2] Department of Chemistry, Langelandsgade 140, 8000 Aarhus C, Denmark., Danmark
  • Jan Johannes Enghild
  • Jørgen Kjems
  • Thomas Tørring
  • Kurt V Gothelf

DNA-protein conjugates are important in bioanalytical chemistry, molecular diagnostics and bionanotechnology, as the DNA provides a unique handle to identify, functionalize or otherwise manipulate proteins. To maintain protein activity, conjugation of a single DNA handle to a specific location on the protein is often needed. However, preparing such high-quality site-specific conjugates often requires genetically engineered proteins, which is a laborious and technically challenging approach. Here we demonstrate a simpler method to create site-selective DNA-protein conjugates. Using a guiding DNA strand modified with a metal-binding functionality, we directed a second DNA strand to the vicinity of a metal-binding site of His6-tagged or wild-type metal-binding proteins, such as serotransferrin, where it subsequently reacted with lysine residues at that site. This method, DNA-templated protein conjugation, facilitates the production of site-selective protein conjugates, and also conjugation to IgG1 antibodies via a histidine cluster in the constant domain.

OriginalsprogEngelsk
TidsskriftNature Chemistry
Vol/bind6
Nummer9
Sider (fra-til)804-809
Antal sider6
ISSN1755-4330
DOI
StatusUdgivet - sep. 2014

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