title: Template-directed covalent conjugation of DNA to native antibodies, transferrin and other metal-binding proteins.
abstract: 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.
authors.affiliation: 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].
authors.forename: Christian B
authors.initials: CB
authors.lastname: Rosen
authors.affiliation: 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].
authors.forename: Anne L B
authors.initials: AL
authors.lastname: Kodal
authors.affiliation: 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.
authors.forename: Jesper S
authors.initials: JS
authors.lastname: Nielsen
authors.affiliation: 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.
authors.forename: David H
authors.initials: DH
authors.lastname: Schaffert
authors.affiliation: Department of Molecular Biology and Genetics, Science Park, Aarhus University, Gustav Wieds Vej 10C, 8000 Aarhus C, Denmark.
authors.forename: Carsten
authors.initials: C
authors.lastname: Scavenius
authors.affiliation: 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.
authors.forename: Anders H
authors.initials: AH
authors.lastname: Okholm
authors.affiliation: 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.
authors.forename: Niels V
authors.initials: NV
authors.lastname: Voigt
authors.affiliation: Department of Molecular Biology and Genetics, Science Park, Aarhus University, Gustav Wieds Vej 10C, 8000 Aarhus C, Denmark.
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Enghild
authors.affiliation: 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.
authors.forename: Jørgen
authors.initials: J
authors.lastname: Kjems
authors.affiliation: 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.
authors.forename: Thomas
authors.initials: T
authors.lastname: Tørring
authors.affiliation: 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.
authors.forename: Kurt V
authors.initials: KV
authors.lastname: Gothelf
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D049452:Green Fluorescent Proteins; D006639:Histidine / Q000737:chemistry; D006801:Humans; D008670:Metals / Q000378:metabolism*; D008969:Molecular Sequence Data; D014168:Transferrin / Q000737:chemistry*
chemical_list: D000906:Antibodies; D002352:Carrier Proteins; D008670:Metals; D014168:Transferrin; D049452:Green Fluorescent Proteins; D006639:Histidine; D004247:DNA