Utjecaj hidrolizata proteina industrijske konoplje na rast, produktivnost i metabolizam CHO DP-12 stanične linije

master's thesis

• Title (english): The impact of hempseed protein hydrolysate on growth, productivity, and metabolism of CHO DP-12 cell line
• Author: Anja Damjanović
• Mentor: Igor Slivac (mentor)
• Committee member: Kristina Radošević (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: 2021-09-24, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

CHO stanična linija je najkorišteniji ekspresijski sustav u biotehnološkoj proizvodnji terapeutika. Jedan od parametara optimizacije te proizvodnje je i odabir hranjivog medija za uzgoj čija formulacija ima veliki utjecaj na ponašanje stanica u kulturi. 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. Tijekom ovog rada, ispitano je djelovanje proteina izoliranih iz industrijske konoplje na stanični rast i produktivnost. Stoga su, hranjivom mediju za uzgoj dodavani nefrakcionirani hidrolizati i frakcije proteina veličina ispod 10 i 1 kDa dobiveni pomoću tri mikrobne proteaze: Alcalase, Neutrase i Protamex u dvije različite koncentracije. Istraživanje je pokazalo da nefrakcionirani hidrolizati dobiveni alkalazom u većoj koncentraciji djeluju citostatično na stanice, dok frakcije proteina dobivenih hidrolizom pomoću neutraze i protamexa mogu imati pozitivan utjecaj na rast i produktivnost CHO DP-12 stanica.

Abstract (english)

CHO cells represent the most frequently used mammalian production host when it comes to the production of therapeutics in biotechnology. One of the optimisation parameters of that production is media formulation which has a major impact on cell behavior in culture. Since media supplementation with animal-derived serum does not fall upon demands of good manufacturing practice, plant protein hydrolysates are being considered as an alternative media supplement. During the study, the impact of proteins isolated from industrial hempseed on cell growth and productivity was evaluated. Therefore, media was supplemented with crude hydrolysate as well as protein fractions of low molecular weight (<10 and <1 kDa) obtained by enzymatic hydrolysis with three different proteases (Alcalase, Neutrase, Protamex) in two different concentrations. Research has shown that crude hydrolysate gained from alcalase in higher concentrations act as a cytostatic while protein fractions attained with hydrolysis using neutrase and protamex can have a positive impact on the growth and productivity of CHO DP-12 cells.

• Keywords: brašno uljne pogače konoplje; CHO DP-12 stanična linija; protutijelo IgG; proteinski hidrolizati
• Keywords (english): hampseed cake; CHO DP-12 cell line; IgG antibody; protein hydrolysate
• Language: croatian
• URN:NBN: urn:nbn:hr:159:980547
• 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: 2021-10-27 11:24:57