The typical primary recovery steps in an E. coli-based bioprocess are to break the cells, efficiently remove the biomass and concentrate the target protein for further chromatographic purification (if needed). High-level production of soluble proteins in the periplasmic space of Pseudomonas fluorescens has enabled the use of efficient non-disruptive methods for the release of the target to the medium. This results in a significantly purer extract and has the potential to eliminate a chromatographic step. We are engaged in the development of a tool-box of periplasmic release approaches, and small-scale techniques to rapidly evaluate the release steps in our Pfçnex Expression TechnologyTM platform. Recently, some of these approaches in small scale evaluation/optimization as well as pilot scale implementation of periplasmic release have been successfully applied towards developing processes for biopharmaceutical proteins.