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Impact of added amino acids on CHO cell productivity during the fed batch bioprocess
Horvat, Jernej (Author), Narat, Mojca (Mentor) More about this mentor... This link opens in a new window, Spadiut, Oliver (Co-mentor)

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Abstract
Over the past 30 years, Chinese hamster ovary (CHO) cells have become a major tool for recombinant protein production, mainly for monoclonal antibodies (mAb) and other biologics. These drugs are widely used to treat cancer, autoimmune disorders and many other diseases, which were considered as untreatable just a few years ago. CHO cells are mammalian cells, which synthesize recombinant products in a form that is suitable for clinical trials and therapies. Nowadays most bioprocesses are fed-batch processes where chemically defined solutions are used to supplement substrates. Such solutions consist of glucose, amino acids, microelements, etc. Despite the addition of the aforementioned solutions during bioprocessing, some of the amino acids are still depleted. In preliminary processes we observed the depletion of asparagine, aspartate, histidine, tryptophan, phenylalanine, glutamate, serine, threonine, glycine, cysteine and lysine. With the adaptation of an already existing feed and the usage of newly developed feeds we were able to increase cell viability and mAb production. These observations showed the importance of understanding the metabolism of CHO cells and the availability of amino acids in the cell culture.

Language:English
Keywords:Amino acids, lactate, CHO cell, feed development, monoclonal antibody production
Work type:Doctoral dissertation (mb31)
Tipology:2.08 - Doctoral Dissertation
Organization:BF - Biotechnical Faculty
Year:2021
Publisher:[J. Horvat]
UDC:60:616-097.3:602.42-7:604.2:661.745(043.3)
COBISS.SI-ID:54007043 This link opens in a new window
Views:386
Downloads:285
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Secondary language

Language:Slovenian
Title:Vpliv dodanih aminokislin na produktivnost celic CHO v bioprocesu z dohranjevanjem
Abstract:
Celice ovarijev kitajskega hrčka (CHO) so v zadnjih letih postale vodilni ekspresijski sistem na področju razvoja ter proizvodnje rekombinantnih proteinov. Monoklonska protitelesa in ostala novodobna zdravila so v redni uporabi za terapijo rakavih, avtoimunskih in drugih bolezni, ki so še pred kratkim veljale za neozdravljive. Celice CHO so zmožne sinteze rekombinantnih proteinov v obliki, primerni za uporabo tako v kliničnih raziskavah kot pozneje v terapijah. V razvoju bioloških zdravil se večinoma uporablja šaržni bioproces z dohranjevanjem, kjer kemijsko definirana gojišča in hranila vsebujejo substrate, kot so glukoza, aminokisline, mikroelemente in druge komponente. Kljub temu, da hranila vsebujejo večino aminokislin, se mnogokrat po končanem bioprocesu izkaže, da jih celice porabijo. V preliminarnih poskusih smo opazili pomanjkanje asparagina, aspartata, histidina, triptofana, fenilalanina, glutamata, serina, treonina, glicina, cisteina in lizina. Z adaptacijo obstoječih in uporabo novih hranil smo povečali celično viabilnost in sintezo monoklonskega protitelesa. S tem smo pokazali, da imata lahko poznavanje celičnega metabolizma in prisotnost aminokislin ključno vlogo pri izboljšanju učinkovitosti celic.

Keywords:Aminokisline, laktat, celice CHO, razvoj hranil, sinteza monoklonskih protiteles

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