Escherichia coli–based Nanobody Production: High-Cell-Density Process Development and Benchmarking of Strain-Plasmid Combinations
This study establishes a robust high-cell-density fed-batch process in *E. coli* that, through the optimization of induction timing, temperature, and host-strain combinations, achieves yields exceeding 2.5 g/L of functional anti-CD45 nanobodies, with periplasmic secretion proving superior to cytoplasmic production in terms of yield, thermal stability, and folding uniformity.