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

Hartmann, Jessica; Münch, Robert C; Freiling, Ruth-Therese; Schneider, Irene C; Dreier, Birgit; Samukange, Washington; Koch, Joachim; Seeger, Markus A; Plückthun, Andreas; Buchholz, Christian J

doi:10.1016/j.omtm.2018.07.001

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Hartmann, J., Münch, R. C., Freiling, R.-T., Schneider, I. C., Dreier, B., Samukange, W., Koch, J., Seeger, M. A., Plückthun, A., & Buchholz, C. J. (2018). A Library-Based Screening Strategy for the Identification of DARPins as Ligands for Receptor-Targeted AAV and Lentiviral Vectors. Molecular Therapy - Methods & Clinical Development, 10, 128–143. https://doi.org/10.1016/j.omtm.2018.07.001

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 biotinylation of the extracellular domains of target receptors as Fc fusion proteins in mammalian cells and the