title: The use of dissolved oxygen-controlled, fed-batch aerobic cultivation for recombinant protein subunit vaccine manufacturing.
abstract: A simple "off-the-shelf" fed-batch approach to aerobic bacterial cultivation for recombinant protein subunit vaccine manufacturing is presented. In this approach, changes in the dissolved oxygen levels are used to adjust the nutrient feed rate (DO-stat), so that the desired dissolved oxygen level is maintained throughout cultivation. This enables high Escherichia coli cell densities and recombinant protein titers. When coupled to a kLa-matched scale-down model, process performance is shown to be consistent at the 2L, 20L, and 200L scales for two recombinant E. coli strains expressing different protein subunit vaccine candidates. Additionally, by mining historical DO-stat nutrient feeding data, a method to transition from DO-stat to a pre-determined feeding profile suitable for larger manufacturing scales without using feedback control is demonstrated at the 2L, 20L, and 200L scales.
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4. Electronic address: patrick.farrell@sanofipasteur.com.
authors.forename: Patrick
authors.initials: P
authors.lastname: Farrell
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Jacob
authors.initials: J
authors.lastname: Sun
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Paul-Philippe
authors.initials: PP
authors.lastname: Champagne
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Heron
authors.initials: H
authors.lastname: Lau
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Meg
authors.initials: M
authors.lastname: Gao
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Hong
authors.initials: H
authors.lastname: Sun
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Arno
authors.initials: A
authors.lastname: Zeiser
authors.affiliation: Bioprocess Research and Development, Sanofi Pasteur, 1755 Steeles Avenue West, Toronto, Canada M2R 3T4.
authors.forename: Tony
authors.initials: T
authors.lastname: D'Amore
mesh_terms: D000332:Aerobiosis; D061249:Batch Cell Culture Techniques / Q000379:methods*; D019149:Bioreactors / Q000382:microbiology; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D010100:Oxygen / Q000032:analysis*; D011994:Recombinant Proteins / Q000378:metabolism*; D013678:Technology, Pharmaceutical / Q000379:methods*; D022223:Vaccines, Subunit / Q000378:metabolism*; D014614:Vaccines, Synthetic / Q000378:metabolism*
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D010100:Oxygen
keywords: Fed-batch cultivation; Recombinant protein; Scale-down