An albumin-oligonucleotide assembly for potential combinatorial drug delivery and half-life extension applications

TY - JOUR

T1 - An albumin-oligonucleotide assembly for potential combinatorial drug delivery and half-life extension applications

AU - Kuhlmann, Matthias

AU - Hamming, Jonas Bohn Refslund

AU - Voldum, Anders

AU - Tsakiridou, Georgia

AU - Larsen, Maja Thim

AU - Schmøkel, Julie

AU - Sohn, Emil

AU - Bienk, Konrad

AU - Schaffert, David Henning

AU - Sørensen, Esben Skipper

AU - Wengel, Jesper

AU - Dupont, Daniel Miotto

AU - Howard, Ken

PY - 2017/12/15

Y1 - 2017/12/15

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 - albumin

KW - anticoagulant

KW - aptamer

KW - covalent conjugation

KW - cysteine 34

KW - drug delivery

KW - factor IXa

KW - half-life extension

KW - neonatal Fc receptor

KW - oligodeoxynucleotides

U2 - 10.1016/j.omtn.2017.10.004

DO - 10.1016/j.omtn.2017.10.004

M3 - Journal article

C2 - 29246307

SN - 2162-2531

VL - 9

SP - 284

EP - 293

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

ER -