High Cell Density Perfusion Culture has a Maintained Exoproteome and Metabolome

QC 20200406
The optimization of bioprocesses for biopharmaceutical manufacturing by Chinese hamster ovary (CHO) cells can be a challenging endeavor and, today, heavily relies on empirical methods treating the bioreactor process and the cells as black boxes. Multi-omics approaches have the potential to reveal otherwise unknown characteristics of these systems and identify culture parameters to more rationally optimize the cultivation process. Here, the authors have applied both metabolomic and proteomic profiling to a perfusion process, using CHO cells for antibody production, to explore how cell biology and reactor environment change as the cell density reaches 200x10(6)cellsmL(-1). The extracellular metabolic composition obtained in perfusion mode shows a markedly more stable profile in comparison to fed-batch, despite a far larger range of viable cell densities in perfusion. This stable profile is confirmed in the extracellular proteosome. Furthermore, the proteomics data shows an increase of structural proteins as cell density increases, which could be due to a higher shear stress and explain the decrease in cell diameter at very high cell densities. Both proteomic and metabolic results shows signs of oxidative stress and changes in glutathione metabolism at very high cell densities. The authors suggest the methodology presented herein to be a powerful tool for optimizing processes of recombinant protein production.

Lundqvist, MagnusKTH,Proteinteknologi,Centre for Advanced BioProduction by Continuous Processing, AdBIOPRO(Swepub:kth)u1hfekeg (author)
Zhang, YeKTH,Wallenberg Center for Protein Research(Swepub:kth)u10tiz4g (author)
Åberg, MagnusStockholms universitet,Institutionen för miljövetenskap och analytisk kemi,Stockholm Univ, Dept Analyt Chem, S-10691 Stockholm, Sweden.(Swepub:su)maber (author)
Edfors, FredrikKTH,Proteinvetenskap,Science for Life Laboratory, SciLifeLab(Swepub:kth)u149ml0e (author)
Bidkhori, GholamrezaKTH,Science for Life Laboratory, SciLifeLab,Skolan för kemi, bioteknologi och hälsa (CBH)(Swepub:kth)u1borjet (author)
Lindahl, AnnaKarolinska Inst, Dept Oncol Pathol, Sci Life Lab, S-17165 Solna, Sweden. (author)
Mie, AxelKarolinska Institutet (author)
Mardinoglu, AdilKTH,Science for Life Laboratory, SciLifeLab,Systembiologi(Swepub:kth)u1t8kmr6 (author)
Field, RaymondMedImmune, Biopharmaceut Dev, Cambridge CB21 6GH, England. (author)
Turner, RichardMedImmune, Biopharmaceut Dev, Cambridge CB21 6GH, England. (author)
Rockberg, JohanKTH,Proteinteknologi,Centre for Advanced BioProduction by Continuous Processing, AdBIOPRO(Swepub:kth)u130jo0a (author)
Chotteau, Véronique,Docent,1963-KTH,Industriell bioteknologi,Centre for Advanced BioProduction by Continuous Processing, AdBIOPRO(Swepub:kth)u1gyk88o (author)
KTHIndustriell bioteknologi (creator_code:org_t)

By the author/editor: Zamani, Leila; Lundqvist, Magnu ...; Zhang, Ye; Åberg, Magnus; Edfors, Fredrik; Bidkhori, Gholam ...; show more...; Lindahl, Anna; Mie, Axel; Mardinoglu, Adil; Field, Raymond; Turner, Richard; Rockberg, Johan; Chotteau, Véroni ...; show less...

About the subject

By the university: Stockholm University; Royal Institute of Technology; Karolinska Institutet

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