Prilagodba uzgoja stanica CHO-DP12 u hranjivom mediju suplementiranom hidrolizatima proteina industrijske konoplje

master's thesis

• Title (english): Adaptation of CHO-DP12 cells in a nutrient medium supplemented with industrial hemp protein hydrolysates
• Author: Lucija Špoljarić
• Mentor: Igor Slivac (mentor)
• Mentor: Marijan Logarušić (pomoć pri izradi, neposredni voditelj)
• Committee member: Višnja Gaurina Srček (predsjednik povjerenstva)
• Committee member: Andreja Leboš-Pavunc (član povjerenstva)
• Committee member: Teuta Murati (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2023-11-02, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

U današnje vrijeme CHO stanična linija je najatraktivniji i najpopularniji ekspresijski sustav u biotehnološkoj proizvodnji terapeutika. Jedan od najbitnijih koraka optimizacije biotehnološke proizvodnje istih je priprema hranjivog medija za uzgoj. Budući da obogaćivanje medija serumom životinjskog podrijetla nije u skladu sa zahtjevima dobre proizvođačke prakse, kao alternativni dodaci mediju intenzivno se istražuju biljni proteinski hidrolizati. Cilj ovog rada bio je ispitivanje djelovanja proteina izoliranih iz industrijske konoplje na rast i produktivnost IgG producirajuće stanične linije CHODP12. Stoga su hranjivom mediju za uzgoj dodavani enzimski hidrolizati navedenih proteina kao i frakcije hidrolizata (<10 kDa) dobivene pomoću mikrobne proteaze, Neutrase. Ranija istraživanja pokazala su uglavnom nepovoljan utjecaj ovih suplemenata na rast stanica, ali i poboljšanu staničnu produktivnost u odnosu na hidrolizatima netretiranu kulturu CHO-DP12. Zato je također ispitano može li početna prilagodba stanica na navedene suplemente u mediju utjecati na kasniji rast i produktivnost stanične kulture, u smislu boljeg rasta i većeg prinosa IgG.

Abstract (english)

In today's time, the CHO cell line is the most attractive and popular expression system in biotechnological production of therapeutics. One of the most crucial steps in optimizing biotechnological production is the preparation of a nutrient medium for cultivation. Since enriching the medium with animal serum is not in line with the requirements of good manufacturing practice, plant protein hydrolysates are actively being researched as alternative additives to the medium. The aim of this study was to investigate the effect of proteins isolated from industrial hemp on the growth and productivity of the IgG-producing cell line CHO-DP12. Therefore, enzymatic hydrolysates of the mentioned proteins as well as hydrolyzate fractions (<10 kDa) obtained using the microbial protease, Neutrase, were added to the nutrient culture medium. Earlier studies showed a mostly unfavorable influence of these supplements on cell growth, but also improved cell productivity compared to the CHO-DP12 culture not treated with hydrolysates. Therefore, it was also examined whether the initial adaptation of the cells to the mentioned supplements in the medium can affect the subsequent growth and productivity of the cell culture, in terms of better growth and higher IgG yield.

• Keywords: CHO DP-12 stanična linija; proteinski hidrolizati; konoplja; protutijelo IgG
• Keywords (english): CHO-DP12 cell line; protein hydrolysates; hemp; IgG antibody
• Language: croatian
• URN:NBN: urn:nbn:hr:159:931477
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-05-23 08:45:53