title: Comprehensive characterization of two different Nicotiana tabacum cell lines leads to doubled GFP and HA protein production by media optimization.
abstract: For over two decades, plant cell cultures have been a promising research platform to express recombinant and therapeutic proteins such as hormones, growth factors, full-size antibodies and antigens. Chosen as a good host for manufacturing recombinant proteins, the Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cell line has been studied in shake flasks by offline analysis of only a few growth parameters. The objective of this study is to comprehensively characterize the growth of a transgenic BY-2 cell line and to investigate the expression profile of the model protein GFP. Based on the correlations between nutrient consumption, cell growth and product formation, the intention is to improve the standard MS-medium. Hereby, multiple growth parameters were analyzed offline and online by using a respiration activity monitoring system (RAMOS). A reproducibly observed shift of the oxygen transfer rate (OTR) could be identified to indicate ammonium depletion in the medium. Concurrent with this ammonium depletion, the total protein concentration began to decrease. After the MS-medium was improved, the GFP concentration nearly doubled. When this improved ammonium enriched medium was applied to another transgenic tobacco cell line similar improvements to the amount of the glycoprotein influenza hemagglutinin (HA) produced by Nicotiana tabacum NT-1 cells could be achieved. Ultimately, this combined offline and online analysis can be successfully used for further cell line characterization and media optimization to improve growth and boost target product formation.
authors: RWTH Aachen University, AVT - Biochemical Engineering, Worringer Weg 1, 52056 Aachen, Germany.
authors: David A
authors: DA
authors: Ullisch
authors: Christina A
authors: CA
authors: Müller
authors: Sabrina
authors: S
authors: Maibaum
authors: Janina
authors: J
authors: Kirchhoff
authors: Andreas
authors: A
authors: Schiermeyer
authors: Stefan
authors: S
authors: Schillberg
authors: Jean L
authors: JL
authors: Roberts
authors: Wiltrud
authors: W
authors: Treffenfeldt
authors: Jochen
authors: J
authors: Büchs
mesh_terms: D003470:Culture Media / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000096:biosynthesis / Q000235:genetics; D030821:Plants, Genetically Modified / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D003470:Culture Media; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D000644:Quaternary Ammonium Compounds; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins