Tuning recombinant protein expression to match secretion capacity

Luminita Gabriela Horga, Samantha Halliwell, Tania Selas Castiñeiras, Chris Wyre, Cristina F. R. O. Matos, Daniela S. Yovcheva, Ross Kent, Rosa Morra, Steven G. Williams, Daniel C. Smith, Neil Dixon

    Research output: Contribution to journalArticlepeer-review


    The secretion of recombinant disulfide-bond containing proteins into the periplasm of Gram-negative bacterial hosts, such as E. coli, has many advantages that can facilitate product isolation, quality and activity. However, the secretion machinery of E. coli has a limited capacity and can become overloaded, leading to cytoplasmic retention of product; which can negatively impact cell viability and biomass accumulation. Fine control over recombinant gene expression offers the potential to avoid this overload by matching expression levels to the host secretion capacity.

    Here we report the application of the RiboTite gene expression control system to achieve this by finely controlling cellular expression levels. The level of control afforded by this system allows cell viability to be maintained, permitting production of high-quality, active product with enhanced volumetric titres.

    The methods and systems reported expand the tools available for the production of disulfide-bond containing proteins, including antibody fragments, in bacterial hosts.
    Original languageEnglish
    JournalMicrobial Cell Factories
    Issue number1
    Early online date22 Dec 2018
    Publication statusE-pub ahead of print - 22 Dec 2018


    • Periplasmic secretion
    • antibody fragments
    • riboswitches
    • codon usage
    • signal peptides


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