title: A high-throughput media design approach for high performance mammalian fed-batch cultures.
abstract: An innovative high-throughput medium development method based on media blending was successfully used to improve the performance of a Chinese hamster ovary fed-batch medium in shaking 96-deepwell plates. Starting from a proprietary chemically-defined medium, 16 formulations testing 43 of 47 components at 3 different levels were designed. Media blending was performed following a custom-made mixture design of experiments considering binary blends, resulting in 376 different blends that were tested during both cell expansion and fed-batch production phases in one single experiment. Three approaches were chosen to provide the best output of the large amount of data obtained. A simple ranking of conditions was first used as a quick approach to select new formulations with promising features. Then, prediction of the best mixes was done to maximize both growth and titer using the Design Expert software. Finally, a multivariate analysis enabled identification of individual potential critical components for further optimization. Applying this high-throughput method on a fed-batch, rather than on a simple batch, process opens new perspectives for medium and feed development that enables identification of an optimized process in a short time frame.
authors.affiliation: Merck Serono SA ; Corsier-sur-Vevey ; Biotech Process Sciences ; Fenil-sur-Corsier, Switzerland.
authors.forename: Yolande
authors.initials: Y
authors.lastname: Rouiller
authors.affiliation: Merck Serono SA ; Corsier-sur-Vevey ; Biotech Process Sciences ; Fenil-sur-Corsier, Switzerland.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Périlleux
authors.affiliation: Merck Serono SA ; Corsier-sur-Vevey ; Biotech Process Sciences ; Fenil-sur-Corsier, Switzerland.
authors.forename: Natacha
authors.initials: N
authors.lastname: Collet
authors.affiliation: Merck Serono SA ; Corsier-sur-Vevey ; Biotech Process Sciences ; Fenil-sur-Corsier, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Jordan
authors.affiliation: Merck Serono SA ; Corsier-sur-Vevey ; Biotech Process Sciences ; Fenil-sur-Corsier, Switzerland.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Stettler
authors.affiliation: Merck Serono SA ; Corsier-sur-Vevey ; Biotech Process Sciences ; Fenil-sur-Corsier, Switzerland.
authors.forename: Hervé
authors.initials: H
authors.lastname: Broly
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods*; D019149:Bioreactors; D016466:CHO Cells; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D005947:Glucose / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D008322:Mammals
chemical_list: D003470:Culture Media; D005947:Glucose
keywords: (micro-scale) cell culture; CHO cells; fed-batch; high-throughput; media blending; medium optimization; monoclonal antibody
references.citation: J Biotechnol. 1999 Apr 15;69(2-3):85-93
references.pmid: 10361720
references.citation: Biochem Eng J. 2001 Mar;7(2):117-119
references.pmid: 11173299
references.citation: Trends Microbiol. 2007 Oct;15(10):469-75
references.pmid: 17920276
references.citation: Glycobiology. 2009 Sep;19(9):936-49
references.pmid: 19494347
references.citation: Cytotechnology. 2013 May;65(3):363-78
references.pmid: 22907508
references.citation: Biotechnol Prog. 2009 Jul-Aug;25(4):980-5
references.pmid: 19533739
references.citation: MAbs. 2010 Sep-Oct;2(5):466-79
references.pmid: 20622510
references.citation: Biotechnol Bioeng. 2009 Aug 15;103(6):1164-75
references.pmid: 19418565
references.citation: Cytotechnology. 2013 Jan;65(1):31-40
references.pmid: 22695856
references.citation: Biotechnol Bioeng. 2009 Aug 15;103(6):1118-28
references.pmid: 19449392
references.citation: Biotechnol Bioeng. 2009 Jan 1;102(1):148-60
references.pmid: 18683260
references.citation: Biotechnol Prog. 2011 Jan-Feb;27(1):2-14
references.pmid: 21312350
references.citation: Cytotechnology. 2005 Jan;47(1-3):37-49
references.pmid: 19003043
references.citation: Biotechnol Prog. 2012 Mar-Apr;28(2):392-405
references.pmid: 22223589
references.citation: Biotechnol Bioeng. 2010 May 1;106(1):57-67
references.pmid: 20073088
references.citation: Biotechnol Bioeng. 2010 Feb 1;105(2):260-75
references.pmid: 19739083
references.citation: Biotechnol Bioeng. 1994 May;43(11):1164-74
references.pmid: 18615530
references.citation: Biotechnol Bioeng. 2012 Jun;109(6):1415-29
references.pmid: 22252269
references.citation: Curr Opin Biotechnol. 2006 Dec;17(6):611-8
references.pmid: 17084609
references.citation: Biotechnol Prog. 2006 Mar-Apr;22(2):595-600
references.pmid: 16599582
references.citation: Biotechnol Bioeng. 2012 Jun;109(6):1404-14
references.pmid: 22407794