title: Parallel experimental design and multivariate analysis provides efficient screening of cell culture media supplements to improve biosimilar product quality.
abstract: Rational and high-throughput optimization of mammalian cell culture media has a great potential to modulate recombinant protein product quality. We present a process design method based on parallel design-of-experiment (DoE) of CHO fed-batch cultures in 96-deepwell plates to modulate monoclonal antibody (mAb) glycosylation using medium supplements. To reduce the risk of losing valuable information in an intricate joint screening, 17 compounds were separated into five different groups, considering their mode of biological action. The concentration ranges of the medium supplements were defined according to information encountered in the literature and in-house experience. The screening experiments produced wide glycosylation pattern ranges. Multivariate analysis including principal component analysis and decision trees was used to select the best performing glycosylation modulators. Subsequent D-optimal quadratic design with four factors (three promising compounds and temperature shift) in shake tubes confirmed the outcome of the selection process and provided a solid basis for sequential process development at a larger scale. The glycosylation profile with respect to the specifications for biosimilarity was greatly improved in shake tube experiments: 75% of the conditions were equally close or closer to the specifications for biosimilarity than the best 25% in 96-deepwell plates. Biotechnol. Bioeng. 2017;114: 1448-1458. © 2017 Wiley Periodicals, Inc.
authors.affiliation: Merck Biopharma, Biotech Process Sciences, Merck Biopharma, Route de Fenil 25, 1804, Corsier-sur-Vevey, Switzerland.|Department of Biotechnology and Biophysics, Biozentrum, Julius-Maximilians-Universität Würzburg, Germany.
authors.forename: David
authors.identifier: 0000-0001-8033-6812
authors.initials: D
authors.lastname: Brühlmann
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Chemical and Bioengineering, ETH Zürich, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Sokolov
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Chemical and Bioengineering, ETH Zürich, Switzerland.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Butté
authors.affiliation: Department of Biotechnology and Biophysics, Biozentrum, Julius-Maximilians-Universität Würzburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Sauer
authors.affiliation: Institute for Biochemical Engineering and Analytics, University of Applied Sciences Giessen, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hemberger
authors.affiliation: Merck Biopharma, Biotech Process Sciences, Merck Biopharma, Route de Fenil 25, 1804, Corsier-sur-Vevey, Switzerland.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Souquet
authors.affiliation: Merck Biopharma, Biotech Process Sciences, Merck Biopharma, Route de Fenil 25, 1804, Corsier-sur-Vevey, Switzerland.
authors.forename: Hervé
authors.initials: H
authors.lastname: Broly
authors.affiliation: Merck Biopharma, Biotech Process Sciences, Merck Biopharma, Route de Fenil 25, 1804, Corsier-sur-Vevey, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Jordan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D061249:Batch Cell Culture Techniques / Q000379:methods* / Q000592:standards; D059451:Biosimilar Pharmaceuticals / Q000378:metabolism* / Q000592:standards; D016466:CHO Cells; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism* / Q000592:standards; D057166:High-Throughput Screening Assays / Q000379:methods* / Q000592:standards; D015999:Multivariate Analysis; D025341:Principal Component Analysis; D015202:Protein Engineering / Q000379:methods / Q000592:standards; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000592:standards; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D059451:Biosimilar Pharmaceuticals; D003470:Culture Media; D011994:Recombinant Proteins
keywords: CHO cell culture; DoE; antibody glycosylation; high-throughput; multivariate analysis; sequential process development