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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Abstract

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.

Original languageEnglish
JournalMolecular Therapy - Nucleic Acids
Volume9
Pages (from-to)284-293
ISSN2162-2531
DOIs
Publication statusPublished - 2017

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Life Expectancy
Half-Life
Albumins
Factor IXa
Pharmaceutical Preparations
Recycling
DNA
Serum
Proteins

Keywords

  • Journal Article

Cite this

Kuhlmann, M., Hamming, J. B. R., Voldum, A., Tsakiridou, G., Larsen, M. T., Schmøkel, J. S., ... 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
Kuhlmann, Matthias ; Hamming, Jonas B R ; Voldum, Anders ; Tsakiridou, Georgia ; Larsen, Maja T ; Schmøkel, Julie S ; Sohn, Emil ; Bienk, Konrad ; Schaffert, David ; Sørensen, Esben S ; Wengel, Jesper ; Dupont, Daniel M ; Howard, Kenneth A. / An Albumin-Oligonucleotide Assembly for Potential Combinatorial Drug Delivery and Half-Life Extension Applications. In: Molecular Therapy - Nucleic Acids. 2017 ; Vol. 9. pp. 284-293.
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Kuhlmann, M, Hamming, JBR, Voldum, A, Tsakiridou, G, Larsen, MT, Schmøkel, JS, Sohn, E, Bienk, K, Schaffert, D, Sørensen, ES, Wengel, J, Dupont, DM & Howard, KA 2017, 'An Albumin-Oligonucleotide Assembly for Potential Combinatorial Drug Delivery and Half-Life Extension Applications', Molecular Therapy - Nucleic Acids, vol. 9, pp. 284-293. https://doi.org/10.1016/j.omtn.2017.10.004

An Albumin-Oligonucleotide Assembly for Potential Combinatorial Drug Delivery and Half-Life Extension Applications. / Kuhlmann, Matthias; Hamming, Jonas B R; Voldum, Anders; Tsakiridou, Georgia; Larsen, Maja T; Schmøkel, Julie S; Sohn, Emil; Bienk, Konrad; Schaffert, David; Sørensen, Esben S; Wengel, Jesper; Dupont, Daniel M; Howard, Kenneth A.

In: Molecular Therapy - Nucleic Acids, Vol. 9, 2017, p. 284-293.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - An Albumin-Oligonucleotide Assembly for Potential Combinatorial Drug Delivery and Half-Life Extension Applications

AU - Kuhlmann, Matthias

AU - Hamming, Jonas B R

AU - Voldum, Anders

AU - Tsakiridou, Georgia

AU - Larsen, Maja T

AU - Schmøkel, Julie S

AU - Sohn, Emil

AU - Bienk, Konrad

AU - Schaffert, David

AU - Sørensen, Esben S

AU - Wengel, Jesper

AU - Dupont, Daniel M

AU - Howard, Kenneth A

N1 - Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

KW - Journal Article

U2 - 10.1016/j.omtn.2017.10.004

DO - 10.1016/j.omtn.2017.10.004

M3 - Journal article

C2 - 29246307

VL - 9

SP - 284

EP - 293

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

SN - 2162-2531

ER -