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

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 ca

SUBMITTER: Buzas D 

PROVIDER: S-EPMC7616808 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature