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Picomolar SARS-CoV-2 Neutralization Using Multi-Arm PEG Nanobody Constructs.

Ainhoa Moliner-Morro1, Daniel J Sheward1,2, Vivien Karl1

  • 1Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 17177 Stockholm, Sweden.

Biomolecules
|December 16, 2020
PubMed
Summary
This summary is machine-generated.

We developed a fast method to create multivalent nanobody constructs using click chemistry. A tetrameric nanobody construct targeting SARS-CoV-2 showed significantly enhanced neutralization capabilities.

Keywords:
PEG linkerSARS-CoV-2Single-domain antibody fragmentclick chemistrymultivalentnanobodyneutralizationsortase A

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Area of Science:

  • Biotechnology
  • Immunology
  • Molecular Biology

Background:

  • Multivalent antibody constructs are valuable in clinical and biotechnological applications.
  • Nanobodies offer increased valency with a small molecular size, making them ideal for multivalent designs.

Purpose of the Study:

  • To develop a novel, rapid method for generating oriented bi- and multivalent nanobody constructs.
  • To functionalize nanobodies with click chemistry moieties for site-specific conjugation.

Main Methods:

  • Utilized copper-free strain-promoted azide-alkyne click chemistry (SPAAC) for oriented assembly.
  • Employed sortase A for site-specific ligation of click chemistry functional groups to nanobody C-termini.
  • Constructed C-to-C-terminal nanobody fusions and 4-arm polyethylene glycol (PEG) tetrameric nanobody constructs.

Main Results:

  • Successfully generated bi- and multivalent nanobody constructs using the described method.
  • Demonstrated the approach using the SARS-CoV-2 neutralizing nanobody Ty1.
  • A 4-arm PEG-based tetrameric Ty1 construct exhibited dramatically enhanced neutralization of SARS-CoV-2, achieving an IC50 in the low picomolar range.

Conclusions:

  • The developed SPAAC-based method provides a rapid and efficient route for creating oriented multivalent nanobody constructs.
  • Tetrameric nanobody constructs, particularly those based on PEG scaffolds, can significantly potentiate neutralization efficacy against viral targets like SARS-CoV-2.