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Effect of amino acid balance inculture medium on CHO cellmetabolism
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH).
2018 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [sv]

CHO-celler (kinesisk hamster-ovarie) har blivit arbetshorse för industriell produktion av biologiska ämnen. Dekryptering av den inre metabolismen av dessa celler kommer att vara en värdefull information för att designa nytt medium och uppnå avancerad processkontroll. I denna studieobserverades observationerna avseende aminosyrametabolism av CHO-celler som svar på varierad aminosyrabalans i mediet och registrerades. Pseudo-perfusionsmetoden bildade grunden vid utförandet av experimenten för att bestämma upptagnings- och produktionshastigheterna för alla aminosyror i olika odlingsmedium tillsammans med andra viktiga cellkulturmetaboliter såsom glukos, ammoniak, laktat, produkttiter, biomassa och livskraft etc. Celler visade olika metaboliska reaktioner på olika testade balanser av aminosyror. Det genererade svaret registrerades för att använda det med matematiska verktyg i framtiden. Ovannämnda mål uppnåddes genom attgenomföra en serie av 3 experiment. Experiment 01; för att bestämma hur lågt ett kommersiellt medium kan spädas för att fortfarande ha acceptabel tillväxt och prestanda vad gäller produktivitet,lönsamhet etc. Experiment 02; att registrera respons av celler till varierad aminosyrabalansi mediet. Detta experiment redesignades senare för att förstärka det metaboliska svaret som ses iexperimentet. Genomförande av ett liknande experiment vid högre celltäthet trodde att vi kunde se förstärkt metaboliskt svar och detta gjordes som Experiment 03. Resultaten av detta experiment mellan aminosyraupptagning och produktionshastigheter ska ännu inte utföras för att se om vi erhöll en amplifierad svar.

Abstract [en]

CHO (Chinese hamster ovary) cells have become the workhorse for industrial production of biologics.Deciphering the internal metabolism of these cells will be a valuable information to design new medium and achieve advanced process control. In this study, the observations pertaining toamino acid metabolism of CHO cells in response to varied amino acid balance in the medium wasobserved and recorded. Pseudo perfusion method formed the basis in conducting the experimentsto determine the uptake and production rates of all amino acids in different culture medium together with other important cell culture metabolites like glucose, ammonia, lactate, product titer,biomass and viability, etc. Cells showed different metabolic response to different tested balanceof amino acids. The response generated were recorded for using it with mathematical tools infuture. The above objective was achieved by conducting a series of 3 experiments. Experiment 01;to determine how low a commercial medium can be diluted to still have acceptable growth and performance in terms of productivity, viability, etc. Experiment 02; to record response of cells tovaried amino acid balance in the medium. This experiment was later redesigned to amplify themetabolic response seen in the experiment. Conducting a similar experiment at higher cell density was thought to help us see amplified metabolic response and this was done as Experiment 03. Theresults of this experiment in terms of amino acid uptake and production rates is yet to be performedto see if we obtained an amplified response.

Place, publisher, year, edition, pages
2018.
Keywords [en]
CHO cell, small scale systems, amino acid metabolism, pseudo perfusion, mathematical modeling, medium design, advanced process control.
Keywords [sv]
CHO-cell, småskaliga system, aminosyremetabolism, pseudoperfusion, matematisk modellering, medium design, avancerad processkontroll
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:kth:diva-238619OAI: oai:DiVA.org:kth-238619DiVA, id: diva2:1260917
Available from: 2018-11-05 Created: 2018-11-05 Last updated: 2018-11-05Bibliographically approved

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