An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation

Anders Dinesen, Veronica L Andersen, Marwa Elkhashab, Diego Pilati, Pernille Bech, Elisabeth Fuchs, Torbjørn R Samuelsen, Alexander Winther, Yunpeng Cai, Anders Märcher, Archie Wall, Marjan Omer, Jesper S Nielsen, Vijay Chudasama, James R Baker, Kurt V Gothelf, Jesper Wengel, Jørgen Kjems, Kenneth A Howard*

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Combinatorial properties such as long-circulation and site- and cell-specific engagement need to be built into the design of advanced drug delivery systems to maximize drug payload efficacy. This work introduces a four-stranded oligonucleotide Holliday Junction (HJ) motif bearing functional moieties covalently conjugated to recombinant human albumin (rHA) to give a "plug-and-play" rHA-HJ multifunctional biomolecular assembly with extended circulation. Electrophoretic gel-shift assays show successful functionalization and purity of the individual high-performance liquid chromatography-purified modules as well as efficient assembly of the rHA-HJ construct. Inclusion of an epidermal growth factor receptor (EGFR)-targeting nanobody module facilitates specific binding to EGFR-expressing cells resulting in approximately 150-fold increased fluorescence intensity determined by flow cytometric analysis compared to assemblies absent of nanobody inclusion. A cellular recycling assay demonstrated retained albumin-neonatal Fc receptor (FcRn) binding affinity and accompanying FcRn-driven cellular recycling. This translated to a 4-fold circulatory half-life extension (2.2 and 0.55 h, for the rHA-HJ and HJ, respectively) in a double transgenic humanized FcRn/albumin mouse. This work introduces a novel biomolecular albumin-nucleic acid construct with extended circulatory half-life and programmable multifunctionality due to its modular design.

Original languageEnglish
JournalBioconjugate Chemistry
Volume35
Issue2
Pages (from-to)214-222
Number of pages9
ISSN1043-1802
DOIs
Publication statusPublished - Feb 2024

Keywords

  • Animals
  • DNA, Cruciform
  • ErbB Receptors/metabolism
  • Half-Life
  • Humans
  • Infant, Newborn
  • Mice
  • Mice, Transgenic
  • Serum Albumin, Human/metabolism

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