Testing a perfusion small-scale model in ambr 15 and tubespin bioreactors for CHO cell clone comparison

Bokalj, Bor (author), Narat, Mojca (mentor), Svetina, Monika (co-mentor)

Small scale bioreactor systems are important in bioprocess development. They allow much faster bioprocess development, reduce development costs, and ultimately contribute significantly to the biopharmaceuticals developed. Automation is a key step in high-throughput systems because it can reduce the need for human intervention, as well as measurement and bioprocess errors. The thesis compares two small scale models (SSM) with one another in a clone comparison study. Fed batch SSMs were only used for the clone comparison, since they were previously evaluated in a separate study. Two separate clone groups, which were prepared through different procedures, were used for the clone comparison. After comparing all SSMs with one another, we observed that the ambr15 perfusion mimic bioprocess needs to be optimised further to achieve full implementation into a bioprocess development pipeline for high-density perfusion bioprocesses (HDPB). Viability and viable cell density of the CHO culture was not yet comparable to larger scale bioprocesses and TubeSpin system tests. During the clone comparison, we observed that both clone groups performed similarly in TubeSpins. On the other hand, when comparing separate clones, the best clone from Group 1 had a slight advantage in fed batch bioprocesses when it comes to IgG production, while the best clone from Group 2 had a slight advantage in HDPB IgG production. This was observed after comparing the best performing clones from each group and the bioprocess type. The top clone from clone group 2 achieved higher IgG concentration during the bioprocesses after 10 days of incubation. The same was true for the top clone from Group 1 in FB, where the best clone from Group 1 achieved higher IgG concentrations. In the near future, IgG quality will have to be evaluated in each of the bioprocess models in order to establish which CHO clones are the superior ones.

• Language: Slovenian
• Keywords: bioprocesses, small scale, chinese hamster, ovary cells, clone comparison, perfusion, fed batch
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Year: 2023
• PID: 20.500.12556/RUL-150191
• COBISS.SI-ID: 164634371
• Publication date in RUL: 15.09.2023

Secondary language

• Language: English
• Title: Testiranje modelov perfuzijskega gojenja v bioreaktorjih ambr 15 in tubespin za primerjavo klonov celic CHO

Abstract

Modelni bioreaktorski sistemi v manjšem merilu so pomembni pri razvoju bioprocesov za biofarmacevtske učinkovine. Omogočajo visokozmogljiv pospešen razvoj ter zmanjšujejo stroške v razvoju bioprocesov. Avtomatizacija je ključna pri visokozmogljivih sistemih, saj zmanjšuje potrebo po ročnem delu in zmanjšuje napake pri meritvah. V magistrskem delu smo primerjali dva modelna bioreaktorska sistema v manjšem merilu za vrednotenje in izbor klonov CHO celic. Šaržna procesa v malem merilu smo uporabili samo za primerjavo klonov, saj sta bila prej ocenjena kot zanesljiva. Za primerjavo klonov smo uporabili dve skupini klonov, ki sta bili pripravljeni na različne načine. Po primerjavi obeh modelov perfuzije smo ugotovili da je potrebno ambr15 perfuzijski model še dodatno optimizirati, da ga vključimo v razvojni postopek perfuzijskih bioprocesov. Živost in gostota živih celic sta bili prenizki, da bi bile primerljive z bioprocesi večjega obsega in poskusi v centrifugirkah. Pri primerjavi klonov sta obe skupini klonov v centrifugirkah dali podobne rezultate, ko smo primerjali vrednosti obeh skupin klonov. Pri primerjavi posameznih klonov pa je klon skupine ena imel rahlo prednost v šaržnih procesih, klon skupine dve pa je imel rahlo prednost v sistemu s perfuzijo. To smo opazili po primerjavi najboljšega klona iz vsake skupine in vrsto bioprocesa med seboj. Najboljši klon skupine dve je imel boljšo proizvodnjo IgG po 10. dnevu inkubacije. Enako velja za klon skupine ena v šaržnem bioprocesu z dohranjevanjem, kjer je najboljši klon skupine ena imel višjo proizvodnjo IgG pri vsaki meritvi. Za prihodnje poskuse je treba oceniti kakovost protiteles, s čimer bomo prišli do končne ocene katera skupina klonov je dejansko boljša v vsakem od načinov vodenja bioprocesa.

• Keywords: bioprocesi, malo merilo, celice ovarija, kitajski hrček, primerjava klonov, perfuzija, šaržni proces z dohranjevanjem