Evaluation of downstream methods for monoclonal antibodies produced from CHO cell lines

master's thesis

• Title (croatian): Evaluacija metoda za pročišćavanje monoklonalnih protutijela proizvedenih iz CHO stanične linije
• Author: Jakov Cvetković
• Mentor: Igor Slivac (mentor)
• Committee member: Mirela Ivančić Šantek (predsjednik povjerenstva)
• Committee member: Višnja Gaurina Srček (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: 2023-04-04, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

CHO culture is a well-charaterized cell line that has been used for years in the field of biomanufacturing to produce various valuable pharmaceutical components, such as monoclonal antibodies that are used for the treatment of numerous diseases. Novel, more cost-effective downstream techniques and methods are continuously being developed by large companies. This thesis focused on the characterization of downstream equipment, mainly three components: depth filters (used for filtration), hollow fibers (used for concentration), and ion exchange membrane (used for anionic exchange, or AEX). For the specific process of trastuzumab purification, ErtelAlsop B4E7 filters were proven to be effective in the DNA removal step. In the critical flux experiment, the Microza hollow fibers (0.2 μm pores) could sustain the highest transmembrane flux before significant fouling occurred. Lastly, AEX was effective in removing host cell proteins/DNA.

Abstract (croatian)

CHO stanice dobro su definirana kultura koje se koriste u proizvodnji mnogih farmaceutskih komponenata, poput monoklonalnih protutijela koja se koriste za prevenciju mnogih bolesti. U svrhu smanjenja troškova, mnoga velika poduzeća kontinuirano razvijaju nove metode pročišćavanja ovih vrijednih spojeva. Tijekom ovog rada ispitana je oprema za pročišćavanje, uz naglasak na tri glavna tipa: dubinski filteri (korišteni za filtraciju), membrane (korištene za koncentraciju) i membranski adsorberi (korištene za anionsku izmjenu). Za specifičan proces pročišćavanja Trastuzumab antitijela, ErtelAlsop B4E7 filteri su dokazano efektivni. Nadalje, ispitivane Microza membrane (pore veličina 0.2 μm) su izdržale najveći protok bez pojave začepljenja. Na kraju, anionska izmjena pokazala se najefektivnijom pri uklanjanju neželjenih staničnih proteina i DNK.

• Keywords: monoclonal antibodies; downstream processing; filters; hollow fibers; ion exchange
• Keywords (croatian): monoklonalna protutijela; procesi izdvajanja; filteri; membrane; ionska izmjena
• Language: english
• URN:NBN: urn:nbn:hr:159:264444
• Study programme: Title: Bioprocess Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-05-21 13:30:50