Fig. 3

5 Nanobody molecules (D, E, F, G and H) were used to benchmark the methanol-free platform strain co-expressing HAC1 (grey bar). As a reference the 5 Nanobodies were expressed under methanol-free conditions in the wild type P. pastoris (orange bar) and in the methanol-based system co-expressing RPP0, KAR2, PDI1 and HAC1 (blue bar). Cultivations were performed in fed-batch fermentation according to a generic methanol-based or methanol-free cultivation conditions as described in materials and methods. Indicated titers in cell broth are measured using PA-HPLC