Site-Selective Protein-Oligonucleotide Conjugation Kit for Metal Binding Proteins (includes His-tags)
The oligonucleotide-protein conjugation kits allow easy access to site-selectively conjugated oligonucleotide-modified protein conjugates. The oligonucleotides have been modified with the necessary reacting groups for conjugation meaning that no pre-activation steps are required. The conjugation chemistry used provides stable covalent linkages that is cannot be cleaved hydrolytically. Furthermore, the linkage provided between the oligonucleotide and the protein is short placing the oligonucleotide close to the surface of the protein.
The chemistry of the kits are built on the methodology described by Rosen et al1 and Kodal et al2 taking advantage of non-covalent interactions to direct the labeling reaction, thus, providing a site-selective conjugation reaction. In these kits, it is required that a metal complex can be formed with the protein. His-tagged proteins are common, as the tag allows for easy purification of the protein of interest. However, as the His-tag is capable of forming metal complexes, it also for the use of these oligonucleotide-protein conjugation kits. If the protein is inherently capable of forming metal complexes (e.g. Transferrin) a His-tag is not a requirement. Furthermore, IgG1-type antibodies contain a His-cluster in the constant domain allowing the kits to used for labeling of these types of antibodies.
Proteins that can be labled with the Oligonucleotide-Protein Conjugation - AffyKit
- Genetically modified: e.g. His-tagged proteins
- Non-genetically modified proteins: e.g. Transferrin, IgG1-type antibodies
The kits contain modified oligonucleotides and buffers needed to perform the conjugation reaction, however, sodium cyanoborohydride (NaCNBH3) is not included in the kits. Affycon have made three kits, which contain different sequences of the oligonucleotides. Affycon also provides custom tailoring of sequences fitting your needs.Table 1:
NB: The kits do not contain sodium cyanoborohydride (NaCNBH3), which is required for the conjugation reaction.
 Rosen, C.B.; Kodal, A. L. B.; Nielsen, J. S.; Schaffert, D. H.; Scavenius, C.; Okholm, A. H.; Voigt, N. V.; Enghild, J. J.; Kjems, J.; Tørring, T.; Gothelf, K. V. Nat. Chem. 2014, 6, 804-809
 Kodal, A. L. B.; Rosen, C. B.; Mortensen, M. R.; Tørring, T.; Gothelf, K. V. ChemBioChem 2016, 17, 1338-1342