An Albumin-Oligonucleotide Assembly for Potential Combinatorial Drug Delivery and Half-Life Extension Applications

Matthias Kuhlmann, Jonas B R Hamming, Anders Voldum, Georgia Tsakiridou, Maja T Larsen, Julie S Schmøkel, Emil Sohn, Konrad Bienk, David Schaffert, Esben S Sørensen, Jesper Wengel, Daniel M Dupont, Kenneth A Howard

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Abstrakt

The long blood circulatory property of human serum albumin, due to engagement with the cellular recycling neonatal Fc receptor (FcRn), is an attractive drug half-life extension enabling technology. This work describes a novel site-specific albumin double-stranded (ds) DNA assembly approach, in which the 3' or 5' end maleimide-derivatized oligodeoxynucleotides are conjugated to albumin cysteine at position 34 (cys34) and annealed with complementary strands to allow single site-specific protein modification with functionalized ds oligodeoxynucleotides. Electrophoretic gel shift assays demonstrated successful annealing of complementary strands bearing Atto488, 6-carboxyfluorescein (6-FAM), or a factor IXa aptamer to the albumin-oligodeoxynucleotide conjugate. A fluorometric factor IXa activity assay showed retained aptamer inhibitory activity upon assembly with the albumin and completely blocked factor IXa at a concentration of 100 nM for 2 hr. The assembled construct exhibited stability in serum-containing buffer and FcRn engagement that could be increased using an albumin variant engineered for higher FcRn affinity. This work presents a novel albumin-oligodeoxynucleotide assembly technology platform that offers potential combinatorial drug delivery and half-life extension applications.

OriginalsprogEngelsk
TidsskriftMolecular Therapy - Nucleic Acids
Vol/bind9
Sider (fra-til)284-293
ISSN2162-2531
DOI
StatusUdgivet - 2017

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Kuhlmann, M., Hamming, J. B. R., Voldum, A., Tsakiridou, G., Larsen, M. T., Schmøkel, J. S., Sohn, E., Bienk, K., Schaffert, D., Sørensen, E. S., Wengel, J., Dupont, D. M., & Howard, K. A. (2017). An Albumin-Oligonucleotide Assembly for Potential Combinatorial Drug Delivery and Half-Life Extension Applications. Molecular Therapy - Nucleic Acids, 9, 284-293. https://doi.org/10.1016/j.omtn.2017.10.004