Primerjava bioprocesov s celično linijo CHO v različnih bioreaktorjih

diplomsko delo univerzitetnega študijskega programa I. stopnje

Matic Košir (Author), Urban Bren (Mentor), Matjaž Brinc (Co-mentor)

Abstract

Farmacevtska industrija dandanes stremi k iskanju biološko ustreznejših rešitev in prehodu iz sintetičnih k biološkim produktom. Biofarmacevtiki predstavljajo vzpenjajoči sektor v farmacevtski industriji in so uporabni za širok spekter indikacij, vključujoč onkologijo in revmatologijo. Razvoj in optimizacija bioprocesov na laboratorijski skali je ključnega pomena za poznavanje procesa in prenos v proizvodnjo. Centralni faktor za razvoj in proizvodnjo biološkega zdravila je bioreaktor. S svojo inovativno zasnovo in visokokakovostnimi materiali mora zagotavljati optimalno in stabilno okolje za gojenje in rast celic. Zadnja leta so v porastu bioreaktorji za enkratno uporabo. Različne študije so pokazale njihove prednosti, ki so pripomogle k nadaljnjim razvojem bioloških zdravil. Tako je prišlo do razvoja nove množice bioreaktorjev, ki se razlikujejo predvsem po obliki, vnosu moči, instrumentaciji in volumnu posode za kultivacijo. Namen raziskovalnega dela je bil izvesti 14-dnevni bioproces v 4-litrskem bioreaktorskem sistemu BIOne podjetja Distek in ga primerjati z referenčnima bioprocesoma. Prvi referenčni bioproces je bil izpeljan v mikrobioreaktorskem sistemu ambr® 250, drugi pa v 1000-litrskem bioreaktorju HyClone proizvajalca Thermo Scientific. Vse bioprocese smo izvedli z istim klonom celic in pod enakimi pogoji. Bioprocesne parametre smo spremljali s pomočjo različnih sond in analitskih naprav. Pridobljene meritve smo obdelali z različnimi statističnimi metodami in jih predstavili na diagramih. Osredotočili smo se na kinetiko rasti, metabolizem in produktivnost sesalskih celic. Rezultati kažejo, da je bioreaktorski sistem pri optimalnih pogojih zagotovil uspešno izvedbo bioprocesa s sesalsko celično linijo CHO. Ugotovili smo zadovoljivo primerljivost med bioprocesi. Na podlagi preučevanih koncentracij metabolitov in substratov, osmolalnosti, kinetiko rasti in koncentracijo produkta smo zaključili, da je omenjen bioreaktorski sistem primeren za nadaljnje študije na naši laboratorijski skali. V prihodnosti nameravamo stremeti k eksperimentom z drugimi celičnimi linijami in testiranju različnih parametrov. Uspešna izvedba bi nam prinesla zaupanje v ta bioreaktorski sistem in omogočila prevzem za širšo uporabo.

Keywords

bioreaktorji za enkratno uporabo;monoklonska protitelesa;biofarmacevtika;šaržni bioproces z dohranjevanjem;kinetika in metabolizem CHO celic;diplomske naloge;

Data

Language:	Slovenian
Year of publishing:	2020
Typology:	2.11 - Undergraduate Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[M. Košir]
UDC:	602.4:57.086.83(043.2)
COBISS:	33830147
Views:	452
Downloads:	63
Average score:	0 (0 votes)
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Other data

Secondary language:	English
Secondary title:	Comparison of bioprocesses with CHO cell line in different bioreactors
Secondary abstract:	The pharmaceutical industry nowadays is striving to find more biologically appropriate solutions and to switch from synthetic to biologic products. Biopharmaceuticals represent an emerging sector in the pharmaceutical industry and are useful for a wide range of indications, including oncology and rheumatology. The development and optimization of bioprocesses on a laboratory scale is of crucial importance to understand the process and the transfer to production. Bioreactor is the central factor for biological drug development and production. Its innovative design and high-quality materials provide an optimal and stable environment for cell growth and their cultivation. In recent years disposable bioreactors have been on the rise. Various studies have shown their benefits which have contributed to the further development of biological drugs. Thus, a new set of bioreactors has been invented which differs predominantly in shape, power input, instrumentation and in volume of the cultivation vessel. The purpose of this research was to perform a 14-day bioprocess in a 4-liter bioreactor system BIOne (Distek) and compare it with the reference bioprocesses. The first reference bioprocess was performed in a microbioreactor system ambr® 250, and the second in a 1000-liter HyClone bioreactor manufactured by Thermo Scientific. All bioprocesses were performed with the same cell clone and under the same condition. Bioprocess parameters were monitored using various probes and analytical devices. The obtained measurements were analysed with the help of various graphical methods which are further presented in diagrams. Our focus was on cell growth kinetics, metabolism, and on mammalian cell productivity. The results show that under optimal conditions the bioreactor system ensured a successful implementation of the bioprocess with the mammalian CHO cell line. We found a satisfactory comparability between the bioprocesses. It can be concluded that, based on the studied concentrations of metabolites and substrates, osmolality, growth kinetics and product concentration, the aforementioned bioreactor system is suitable for further studies on the laboratory scale. In the future, we intend to experiment with other cell lines, and to test different parameters. A successful implementation would be a guarantee of success of this bioreactor system and it would enable its acceptance for wider use.
Secondary keywords:	disposable bioreactors;monoclonal antibodies;biopharmacy;fed-batch bioprocesses;kinetics and metabolism of CHO cells.;
Type (COBISS):	Bachelor thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	X, 39 str.
ID:	11938497