title: Novel micro-bioreactor high throughput technology for cell culture process development: Reproducibility and scalability assessment of fed-batch CHO cultures.
abstract: With increasing timeline pressures to get therapeutic and vaccine candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for process development. One such high-throughput system is the SimCell platform, a robotically driven, cell culture bioreactor system developed by BioProcessors Corp. This study describes the use of the SimCell micro-bioreactor technology for fed-batch cultivation of a GS-CHO transfectant expressing a model IgG4 monoclonal antibody. Cultivations were conducted in gas-permeable chambers based on a micro-fluidic design, with six micro-bioreactors (MBs) per micro-bioreactor array (MBA). Online, non-invasive measurement of total cell density, pH and dissolved oxygen (DO) was performed. One hundred fourteen parallel MBs (19 MBAs) were employed to examine process reproducibility and scalability at shake flask, 3- and 100-L bioreactor scales. The results of the study demonstrate that the SimCell platform operated under fed-batch conditions could support viable cell concentrations up to least 12 x 10(6) cells/mL. In addition, both intra-MB (MB to MB) as well as intra-MBA (MBA to MBA) culture performance was found to be highly reproducible. The intra-MB and -MBA variability was calculated for each measurement as the coefficient of variation defined as CV (%) = (standard deviation/mean) x 100. The % CV values for most intra-MB and intra-MBA measurements were generally under 10% and the intra-MBA values were slightly lower than those for intra-MB. Cell growth, process parameters, metabolic and protein titer profiles were also compared to those from shake flask, bench-top, and pilot scale bioreactor cultivations and found to be within +/-20% of the historical averages.
authors.affiliation: Vaccine & Biologics Process Development, Merck Research Laboratories, Merck & Co. Inc., West Point, Pennsylvania 19486, USA. amanullah.ashraf@gene.com
authors.forename: Ashraf
authors.initials: A
authors.lastname: Amanullah
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Otero
authors.forename: Mark
authors.initials: M
authors.lastname: Mikola
authors.forename: Amy
authors.initials: A
authors.lastname: Hsu
authors.forename: Jinyou
authors.initials: J
authors.lastname: Zhang
authors.forename: John
authors.initials: J
authors.lastname: Aunins
authors.forename: H Brett
authors.initials: HB
authors.lastname: Schreyer
authors.forename: James A
authors.initials: JA
authors.lastname: Hope
authors.forename: A Peter
authors.initials: AP
authors.lastname: Russo
mesh_terms: D000818:Animals; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D007074:Immunoglobulin G / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D015203:Reproducibility of Results
chemical_list: D007074:Immunoglobulin G; D011994:Recombinant Proteins