A Library-Based Screening Strategy for the Identification of DARPins as Ligands for Receptor-Targeted AAV and Lentiviral Vectors.

ABSTRACT: Delivering genes selectively to the therapeutically relevant cell type is among the prime goals of vector development. Here, we present a high-throughput selection and screening process that identifies designed ankyrin repeat proteins (DARPins) optimally suited for receptor-targeted gene delivery using adeno-associated viral (AAV) and lentiviral (LV) vectors. In particular, the process includes expression, purification, and in situ biotinylation of the extracellular domains of target receptors as Fc fusion proteins in mammalian cells and the selection of high-affinity binders by ribosome display from DARPin libraries each covering more than 10¹² variants. This way, DARPins specific for the glutamate receptor subunit GluA4, the endothelial surface marker CD105, and the nat

SUBMITTER: Hartmann J 

PROVIDER: S-EPMC6077149 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature