Claudia D'Ercole - Secondment from UNG to RWTH

As part of the EXPAND EV project, Claudia D’Ercole from the University of Nova Gorica completed a three-month secondment at RWTH Aachen University. During this period, she focused on optimizing the expression of two recombinant proteins (a nanobody against CD30 and the extracellular domain of PD-1) using a cell-free expression system. Protein expression was evaluated in different cellular compartments (microsomes and cytosol), selected according to the post-translational modifications required for correct protein folding.

In parallel, the same proteins were expressed in Nicotiana benthamiana to enable further characterization. The anti-CD30 nanobody was analyzed by surface plasmon resonance (SPR) to assess its binding affinity to the CD30 ligand, while PD-1 was characterized with respect to its glycosylation pattern.

In addition, extracellular vesicles (exosomes) were isolated from Nicotiana benthamiana leaves using three different approaches: differential centrifugation, size-exclusion chromatography, and sucrose density gradient centrifugation.

Overall, the secondment provided strong expertise in protein expression across multiple systems and represented a starting point for the optimization of exosome isolation protocols, with the aim of identifying the most efficient and reproducible method.