Escherichia coli is one of the most popular expression systems for the production of recombinant proteins. In the present study, a brief protocol to apply a fed-batch mode at lab scale with E.coli and the scale-up to 30 liters was performed. The high cell density of E.coli growth was achieved using a fed-batch mode in a Bionet F1 Benchtop bioreactor, after characterization in batch mode by using an online optical density probe to register automatically the growth. A specific growth rate of 0.46h-1 was obtained, resulting in a 7-fold biomass concentration yield with the fed-batch strategy, in comparison with the alternative batch mode. The process developed at the bench-top scale was scaled up with a factor of 10, under constant volumetric oxygen transfer coefficients KLa criterion in a Bionet F2-30 bioreactor at pilot scale.