Engineering the ADDobody protein scaffold for generation of high-avidity ADDomer super-binders

Dora Buzas, Huan Sun, Christine Toelzer, Sathish K N Yadav, Ufuk Borucu, Gunjan Gautam, Kapil Gupta, Josh C Bufton, Julien M Capin, Richard B Sessions, Frederic Garzoni, Imre Berger, Christiane H Berger-Schaffitzel

Research output: Contribution to journalArticle (Academic Journal)peer-review

Abstract

Adenovirus-derived nanoparticles (ADDomer) comprise 60 copies of adenovirus penton base protein (PBP). ADDomer is thermostable, rendering the storage, transport, and deployment of ADDomer-based therapeutics independent of a cold chain. To expand the scope of ADDomers for new applications, we engineered ADDobodies, representing PBP crown domain, genetically separated from PBP multimerization domain. We inserted heterologous sequences into hyper-variable loops, resulting in monomeric, thermostable ADDobodies expressed at high yields in Escherichia coli. The X-ray structure of an ADDobody prototype validated our design. ADDobodies can be used in ribosome display experiments to select a specific binder against a target, with an enrichment factor of ∼104-fold per round. ADDobodies can be re-converted into ADDomers by genetically reconnecting the selected ADDobody with the PBP multimerization domain from a different species, giving rise to a multivalent nanoparticle, called Chimera, confirmed by a 2.2 Å electron cryo-microscopy structure. Chimera comprises 60 binding sites, resulting in ultra-high, picomolar avidity to the target.
Original languageEnglish
Pages (from-to)342-351.e6
JournalStructure
Volume32
Issue number3
DOIs
Publication statusPublished - 9 Jan 2024

Bibliographical note

Publisher Copyright:
© 2023 The Author(s)

Structured keywords

  • Max Planck Bristol
  • Bristol BioDesign Institute

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