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

doi:10.1021/acs.bioconjchem.3c00491

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 af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

2 Citationer (Scopus)

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.

Originalsprog	Engelsk
Tidsskrift	Bioconjugate Chemistry
Vol/bind	35
Nummer	2
Sider (fra-til)	214-222
Antal sider	9
ISSN	1043-1802
DOI	https://doi.org/10.1021/acs.bioconjchem.3c00491
Status	Udgivet - feb. 2024

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Dinesen, A., Andersen, V. L., Elkhashab, M., Pilati, D., Bech, P., Fuchs, E., Samuelsen, T. R., Winther, A., Cai, Y., Märcher, A., Wall, A., Omer, M., Nielsen, J. S., Chudasama, V., Baker, J. R., Gothelf, K. V., Wengel, J., Kjems, J., & Howard, K. A. (2024). An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation. Bioconjugate Chemistry, 35(2), 214-222. https://doi.org/10.1021/acs.bioconjchem.3c00491

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title = "An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation",

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.",

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

author = "Anders Dinesen and Andersen, {Veronica L} and Marwa Elkhashab and Diego Pilati and Pernille Bech and Elisabeth Fuchs and Samuelsen, {Torbj{\o}rn R} and Alexander Winther and Yunpeng Cai and Anders M{\"a}rcher and Archie Wall and Marjan Omer and Nielsen, {Jesper S} and Vijay Chudasama and Baker, {James R} and Gothelf, {Kurt V} and Jesper Wengel and J{\o}rgen Kjems and Howard, {Kenneth A}",

year = "2024",

month = feb,

doi = "10.1021/acs.bioconjchem.3c00491",

language = "English",

volume = "35",

pages = "214--222",

journal = "Bioconjugate Chemistry",

issn = "1043-1802",

publisher = "American Chemical Society",

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Dinesen, A, Andersen, VL, Elkhashab, M, Pilati, D, Bech, P, Fuchs, E, Samuelsen, TR, Winther, A, Cai, Y, Märcher, A, Wall, A, Omer, M, Nielsen, JS, Chudasama, V, Baker, JR, Gothelf, KV, Wengel, J, Kjems, J & Howard, KA 2024, 'An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation', Bioconjugate Chemistry, bind 35, nr. 2, s. 214-222. https://doi.org/10.1021/acs.bioconjchem.3c00491

An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation. / Dinesen, Anders; Andersen, Veronica L; Elkhashab, Marwa et al.
I: Bioconjugate Chemistry, Bind 35, Nr. 2, 02.2024, s. 214-222.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

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T1 - An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation

AU - Dinesen, Anders

AU - Andersen, Veronica L

AU - Elkhashab, Marwa

AU - Pilati, Diego

AU - Bech, Pernille

AU - Fuchs, Elisabeth

AU - Samuelsen, Torbjørn R

AU - Winther, Alexander

AU - Cai, Yunpeng

AU - Märcher, Anders

AU - Wall, Archie

AU - Omer, Marjan

AU - Nielsen, Jesper S

AU - Chudasama, Vijay

AU - Baker, James R

AU - Gothelf, Kurt V

AU - Wengel, Jesper

AU - Kjems, Jørgen

AU - Howard, Kenneth A

PY - 2024/2

Y1 - 2024/2

N2 - 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.

AB - 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.

KW - Animals

KW - DNA, Cruciform

KW - ErbB Receptors/metabolism

KW - Half-Life

KW - Humans

KW - Infant, Newborn

KW - Mice

KW - Mice, Transgenic

KW - Serum Albumin, Human/metabolism

UR - http://www.scopus.com/inward/record.url?scp=85183491100&partnerID=8YFLogxK

U2 - 10.1021/acs.bioconjchem.3c00491

DO - 10.1021/acs.bioconjchem.3c00491

M3 - Journal article

C2 - 38231391

SN - 1043-1802

VL - 35

SP - 214

EP - 222

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 2

ER -

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

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