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Rosen, Christian B; Kodal, Anne L B; Nielsen, Jesper S; Schaffert, David H; Scavenius, Carsten; Okholm, Anders H; Voigt, Niels V; Enghild, Jan J; Kjems, Jørgen; Tørring, Thomas; Gothelf, Kurt V
Nature chemistry, 09/2014, Letnik: 6, Številka: 9Journal Article
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.
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