Plug-and-play nucleic acid-mediated multimerization of biparatopic nanobodies for molecular imaging

Laura Teodori; Sarah Kroer; Marjan Omer; Veronica Liv Andersen; Pernille Bech; Junyi Su; Jessica Bridoux; Jesper Sejrup Nielsen; Mathias Bøjlesen Bertelsen; Sophie  Hernot; Kurt Vesterager Gothelf; Jørgen Kjems

doi:10.1016/j.omtn.2024.102305

Plug-and-play nucleic acid-mediated multimerization of biparatopic nanobodies for molecular imaging

Laura Teodori, Sarah Kroer, Marjan Omer, Veronica Liv Andersen, Pernille Bech, Junyi Su, Jessica Bridoux, Jesper Sejrup Nielsen, Mathias Bøjlesen Bertelsen, Sophie Hernot, Kurt Vesterager Gothelf, Jørgen Kjems^*

^*Corresponding author af dette arbejde

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

2 Citationer (Scopus)

Abstract

In cancer molecular imaging, selecting binders with high specificity and affinity for biomarkers is paramount for achieving high-contrast imaging within clinical time frames. Nanobodies have emerged as potent candidates, surpassing antibodies in pre-clinical imaging due to their convenient production, rapid renal clearance, and deeper tissue penetration. Multimerization of nanobodies is a popular strategy to enhance their affinity and pharmacokinetics; however, traditional methods are laborious and may yield heterogeneous products. In this study, we employ a Holliday junction (HJ)-like nucleic acid-based scaffold to create homogeneous nanostructures with precise multivalent and multiparatopic nanobody displays. The plug-and-play assembly allowed the screening of several nanobody multimer configurations for the detection of the breast cancer biomarker, human epidermal growth factor receptor 2 (HER2). In vitro studies demonstrated significant improvements in binding avidity, particularly with the biparatopic construct exhibiting high sensitivity, surpassing that of traditional antibody-based cell binding. Furthermore, our HJ platform allowed for adaptation from fluorescence-based to nuclear imaging, as demonstrated in xenografted mice, thereby allowing for future in vivo applications. This work highlights the potential of nucleic acid-mediated multimerization to markedly enhance nanobody binding, by exploring synergistic combinations and offering versatility for both in vitro diagnostics and cancer molecular imaging with prospects for future theranostic applications.

Originalsprog	Engelsk
Artikelnummer	102305
Tidsskrift	Molecular Therapy - Nucleic Acids
Vol/bind	35
Nummer	3
Antal sider	14
ISSN	2162-2531
DOI	https://doi.org/10.1016/j.omtn.2024.102305
Status	Udgivet - 10 sep. 2024

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10.1016/j.omtn.2024.102305Licens: CC BY-NC-ND

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title = "Plug-and-play nucleic acid-mediated multimerization of biparatopic nanobodies for molecular imaging",

abstract = "In cancer molecular imaging, selecting binders with high specificity and affinity for biomarkers is paramount for achieving high-contrast imaging within clinical time frames. Nanobodies have emerged as potent candidates, surpassing antibodies in pre-clinical imaging due to their convenient production, rapid renal clearance, and deeper tissue penetration. Multimerization of nanobodies is a popular strategy to enhance their affinity and pharmacokinetics; however, traditional methods are laborious and may yield heterogeneous products. In this study, we employ a Holliday junction (HJ)-like nucleic acid-based scaffold to create homogeneous nanostructures with precise multivalent and multiparatopic nanobody displays. The plug-and-play assembly allowed the screening of several nanobody multimer configurations for the detection of the breast cancer biomarker, human epidermal growth factor receptor 2 (HER2). In vitro studies demonstrated significant improvements in binding avidity, particularly with the biparatopic construct exhibiting high sensitivity, surpassing that of traditional antibody-based cell binding. Furthermore, our HJ platform allowed for adaptation from fluorescence-based to nuclear imaging, as demonstrated in xenografted mice, thereby allowing for future in vivo applications. This work highlights the potential of nucleic acid-mediated multimerization to markedly enhance nanobody binding, by exploring synergistic combinations and offering versatility for both in vitro diagnostics and cancer molecular imaging with prospects for future theranostic applications. ",

keywords = "MT: Oligonucleotides: Diagnostics and Biosensors, biparatopic, breast cancer, molecular imaging, multimerization, nanobodies, nanostructure, nucleic acid, single-domain antibodies, theranostic platform, tumor targeting",

author = "Laura Teodori and Sarah Kroer and Marjan Omer and Andersen, {Veronica Liv} and Pernille Bech and Junyi Su and Jessica Bridoux and Nielsen, {Jesper Sejrup} and Bertelsen, {Mathias B{\o}jlesen} and Sophie Hernot and Gothelf, {Kurt Vesterager} and J{\o}rgen Kjems",

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AU - Teodori, Laura

AU - Kroer, Sarah

AU - Omer, Marjan

AU - Andersen, Veronica Liv

AU - Bech, Pernille

AU - Su, Junyi

AU - Bridoux, Jessica

AU - Nielsen, Jesper Sejrup

AU - Bertelsen, Mathias Bøjlesen

AU - Hernot, Sophie

AU - Gothelf, Kurt Vesterager

AU - Kjems, Jørgen

PY - 2024/9/10

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N2 - In cancer molecular imaging, selecting binders with high specificity and affinity for biomarkers is paramount for achieving high-contrast imaging within clinical time frames. Nanobodies have emerged as potent candidates, surpassing antibodies in pre-clinical imaging due to their convenient production, rapid renal clearance, and deeper tissue penetration. Multimerization of nanobodies is a popular strategy to enhance their affinity and pharmacokinetics; however, traditional methods are laborious and may yield heterogeneous products. In this study, we employ a Holliday junction (HJ)-like nucleic acid-based scaffold to create homogeneous nanostructures with precise multivalent and multiparatopic nanobody displays. The plug-and-play assembly allowed the screening of several nanobody multimer configurations for the detection of the breast cancer biomarker, human epidermal growth factor receptor 2 (HER2). In vitro studies demonstrated significant improvements in binding avidity, particularly with the biparatopic construct exhibiting high sensitivity, surpassing that of traditional antibody-based cell binding. Furthermore, our HJ platform allowed for adaptation from fluorescence-based to nuclear imaging, as demonstrated in xenografted mice, thereby allowing for future in vivo applications. This work highlights the potential of nucleic acid-mediated multimerization to markedly enhance nanobody binding, by exploring synergistic combinations and offering versatility for both in vitro diagnostics and cancer molecular imaging with prospects for future theranostic applications.

AB - In cancer molecular imaging, selecting binders with high specificity and affinity for biomarkers is paramount for achieving high-contrast imaging within clinical time frames. Nanobodies have emerged as potent candidates, surpassing antibodies in pre-clinical imaging due to their convenient production, rapid renal clearance, and deeper tissue penetration. Multimerization of nanobodies is a popular strategy to enhance their affinity and pharmacokinetics; however, traditional methods are laborious and may yield heterogeneous products. In this study, we employ a Holliday junction (HJ)-like nucleic acid-based scaffold to create homogeneous nanostructures with precise multivalent and multiparatopic nanobody displays. The plug-and-play assembly allowed the screening of several nanobody multimer configurations for the detection of the breast cancer biomarker, human epidermal growth factor receptor 2 (HER2). In vitro studies demonstrated significant improvements in binding avidity, particularly with the biparatopic construct exhibiting high sensitivity, surpassing that of traditional antibody-based cell binding. Furthermore, our HJ platform allowed for adaptation from fluorescence-based to nuclear imaging, as demonstrated in xenografted mice, thereby allowing for future in vivo applications. This work highlights the potential of nucleic acid-mediated multimerization to markedly enhance nanobody binding, by exploring synergistic combinations and offering versatility for both in vitro diagnostics and cancer molecular imaging with prospects for future theranostic applications.

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KW - biparatopic

KW - breast cancer

KW - molecular imaging

KW - multimerization

KW - nanobodies

KW - nanostructure

KW - nucleic acid

KW - single-domain antibodies

KW - theranostic platform

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DO - 10.1016/j.omtn.2024.102305

M3 - Journal article

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VL - 35

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

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M1 - 102305

ER -

Plug-and-play nucleic acid-mediated multimerization of biparatopic nanobodies for molecular imaging

Abstract

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