title: A strategy for clone selection under different production conditions.
abstract: Top performing clones have failed at the manufacturing scale while the true best performer may have been rejected early in the screening process. Therefore, the ability to screen multiple clones in complex fed-batch processes using multiple process variations can be used to assess robustness and to identify critical factors. This dynamic ranking of clones' strategy requires the execution of many parallel experiments than traditional approaches. Therefore, this approach is best suited for micro-bioreactor models which can perform hundreds of experiments quickly and efficiently. In this study, a fully monitored and controlled small scale platform was used to screen eight CHO clones producing a recombinant monoclonal antibody across several process variations, including different feeding strategies, temperature shifts and pH control profiles. The first screen utilized 240 micro-bioreactors were run for two weeks for this assessment of the scale-down model as a high-throughput tool for clone evaluation. The richness of the outcome data enable to clearly identify the best and worst clone as well as process in term of maximum monoclonal antibody titer. The follow-up comparison study utilized 180 micro-bioreactors in a full factorial design and a subset of 12 clone/process combinations was selected to be run parallel in duplicate shake flasks. Good correlation between the micro-bioreactor predictions and those made in shake flasks with a Pearson correlation value of 0.94. The results also demonstrate that this micro-scale system can perform clone screening and process optimization for gaining significant titer improvements simultaneously. This dynamic ranking strategy can support better choices of production clones.
authors: Seahorse Bioscience Inc, 16 Esquire Road, Billerica, MA 01862, USA. rlegmann@seahorsebio.com
authors: Rachel
authors: R
authors: Legmann
authors: Brian
authors: B
authors: Benoit
authors: Ronald W
authors: RW
authors: Fedechko
authors: Cynthia L
authors: CL
authors: Deppeler
authors: Sriram
authors: S
authors: Srinivasan
authors: Russell H
authors: RH
authors: Robins
authors: Ellen L
authors: EL
authors: McCormick
authors: David A
authors: DA
authors: Ferrick
authors: Seth T
authors: ST
authors: Rodgers
authors: A Peter
authors: AP
authors: Russo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D002999:Clone Cells; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D008904:Miniaturization; D011994:Recombinant Proteins / Q000096:biosynthesis*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins