Improving monoclonal antibody production from Chinese hamster ovary cells
dc.contributor.advisor
Rosser, Susan
dc.contributor.advisor
Menolascina, Filippo
dc.contributor.author
Donaldson, James
dc.contributor.sponsor
FUJIFILM Diosynth Biotechnologies (FDB)
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dc.contributor.sponsor
IBioIC
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dc.date.accessioned
2024-05-13T10:14:21Z
dc.date.available
2024-05-13T10:14:21Z
dc.date.issued
2024-05-13
dc.description.abstract
Chinese Hamster Ovary (CHO) cells are used for the production of many
therapeutic proteins, including the majority of monoclonal antibodies (mAbs).
While significant progress has been made in improving mAb production
using CHO cells, challenges remain in producing sufficient quantities of nextgeneration
biologics, such as fusion proteins and bi-specific antibodies.
Additionally, production instability (i.e., loss of mAb productivity) during long
term culture remains a significant problem. Stability studies, which take
several months to complete, are a bottleneck in cell line development.
A landing pad system for expression vector component comparison was
integrated into a CHO host cell line. To demonstrate the functionality of the
system, the strength of various constitutive promoters were tested by
quantifying mCherry expression levels. This system will help facilitate the
development of future expression vectors and enable systematic
identification and optimisation of components to enhance CHO cell
productivity.
A system which can predict the production stability of recombinant CHO cell
lines, prior to the completion of a stability study, was envisaged. To pursue
this, the production stabilities of 2 monoclonal CHO cell lines (32-124 and
32-121) producing the same mAb were characterised over ~60 generations.
Loss of volumetric productivity during long term culture was observed in the
32-121 cell line whereas 32-124 exhibited a minimal decrease. Further
analyses of growth characteristics, specific productivity, metabolite profiles,
gene copy number, and transgene mRNA expression were subsequently
conducted to investigate the underlying cause of productivity loss. The
Berkley Lights Beacon® system was subsequently used to compare the two
cell lines at generation 15. The results showed differences in both production
and growth variability between the two cell populations, as well as an
increased frequency of low-producing fast-growing cells in the 32-121 cell
line. The method developed during this study will add to existing strategies
for identifying early indicators of instability in CHO cell lines.
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dc.identifier.uri
https://hdl.handle.net/1842/41771
dc.identifier.uri
http://dx.doi.org/10.7488/era/4494
dc.language.iso
en
en
dc.publisher
The University of Edinburgh
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dc.relation.hasversion
Donaldson, J., Kleinjan, D.-J., Rosser, S., 2022. Synthetic biology approaches for dynamic CHO cell engineering. Current Opinion in Biotechnology 78, 102806.
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dc.relation.hasversion
Donaldson, J.S., Dale, M.P., Rosser, S.J., 2021. Decoupling Growth and Protein Production in CHO Cells: A Targeted Approach. Front. Bioeng. Biotechnol. 9. https://doi.org/10.3389/fbioe.2021.658325
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dc.subject
Chinese hamster
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dc.subject
ovary cells
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dc.subject
monoclonal antibody production
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dc.subject
Chinese Hamster Ovary (CHO) cells
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dc.subject
monoclonal antibodies (mAbs)
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dc.subject
nextgeneration biologics
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dc.subject
mAb productivity
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dc.title
Improving monoclonal antibody production from Chinese hamster ovary cells
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dc.type
Thesis or Dissertation
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dc.type.qualificationlevel
Doctoral
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dc.type.qualificationname
PhD Doctor of Philosophy
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