TY - JOUR
T1 - Plug-and-play nucleic acid-mediated multimerization of biparatopic nanobodies for molecular imaging
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
Y1 - 2024/9/10
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.
U2 - 10.1016/j.omtn.2024.102305
DO - 10.1016/j.omtn.2024.102305
M3 - Journal article
C2 - 39281705
SN - 2162-2531
VL - 35
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
IS - 3
M1 - 102305
ER -