Combinatorial protein engineering for gene therapy and affinity protein applications

The thesis evaluates affinity proteins for selective gene therapies and bioprocessing, using deep sequencing in phage display to identify Affibody and nanobody binders with differing kinetics and target preferences. One paper characterises two nanobodies as protease inhibitors; another expands nanobody family analysis.

For gene delivery, Affibody ligands were integrated into AAV surfaces; the vehicles selectively transduced target-expressing cells in vitro and, in vivo, reduced liver uptake while redirecting transduction. The same binders showed selective in vitro transduction as AAV ligands. Transduction may depend on epitope, kinetics and capsid integration site, so binder diversity matters.

An Affimer ligand for AAV9 chromatography matched commercial resins in recovery and quality, with milder elution. Another paper assesses plasmid purification effects on rAAV production.