The thesis work consists of a list of figures, a list of tables, a list of contents, a list of symbols and acronyms, an abstract, an introduction, a literature review, an experimental part, results and discussions, a conclusion and a list of bibliography used. The materials of the thesis work are presented on 140 pages of printed text, including 26 tables and 60 figures. The thesis work is devoted to the optimization of the glycosylation profile of a therapeutic molecule, monoclonal antibody adalimumab, at the stage of cell line cultivation, and to the study of the monoclonal antibody quality attributes that affect its biological properties. During the practical part, studies were carried out to reveal the influence of the composition of the media, feeds and additives on the glycosylation profile of the therapeutic molecule at the cultivation stage. The dependence of the biological effectiveness of the monoclonal antibody adalimumab on the composition of the glycosylated forms in the molecule was confirmed. There were also developed analytical procedures for determining the charge variants profile by cation exchange chromatography, aggregation products by size-exclusion chromatography, residual host cell DNA by real-time PCR. The procedures for determining the aggregates and residual host cell DNA were validated. The performed scientific research and the results obtained in this work can be taken into account and used as popular solutions to practical problems at the stages of technology development for the production of modern highly effective drugs.
Il lavoro di tesi consiste in una lista di figure, tabelle, contenuti, simboli e acronimi, nonché un sommario, un’introduzione, un articolo letterario, una parte sperimentale, un risultato con annessa discussione, una conclusione e una lista dei riferimenti bibliografici utilizzati. I materiali di questo lavoro di Tesi sono presentati in 140 pagine di testo stampato e includono 26 tabelle e 60 figure. Il lavoro di tesi è finalizzato ad ottimizzare il profilo di glicosilazione di una molecola ad uso terapeutico, l’anticorpo monoclonale adalimumab, allo stadio relativo alla coltura cellulare, nonché a studiare la qualità caratteristiche dell’anticorpo che influenza le proprietà biologiche. Durante la parte pratica, sono stati effettuati studi per rivelare l'influenza della composizione di terreni, mangimi e additivi sul profilo di glicosilazione della molecola terapeutica in fase di coltivazione. È stata confermata la dipendenza dell'efficacia biologica dell'anticorpo monoclonale adalimumab dalla composizione delle forme glicosilate nella molecola. Sono state inoltre sviluppate procedure analitiche per determinare il profilo delle varianti di carica mediante cromatografia a scambio cationico, prodotti di aggregazione mediante cromatografia ad esclusione dimensionale, DNA residuo della cellula ospite mediante PCR in tempo reale. Le procedure per determinare gli aggregati e il DNA residuo della cellula ospite sono state convalidate. La ricerca scientifica svolta e i risultati ottenuti in questo lavoro possono essere presi in considerazione e utilizzati come soluzioni comuni a problemi pratici nelle fasi di sviluppo di tecnologie per la produzione di farmaci moderni altamente efficaci.
Optimization of the monoclonal antibody adalimumab glycosilation profile at the cultivation stage and the study of quality attributes affecting its biological properties
STERIN, ILYA;TVERDOKHLEBOVA, ANNA
2019/2020
Abstract
The thesis work consists of a list of figures, a list of tables, a list of contents, a list of symbols and acronyms, an abstract, an introduction, a literature review, an experimental part, results and discussions, a conclusion and a list of bibliography used. The materials of the thesis work are presented on 140 pages of printed text, including 26 tables and 60 figures. The thesis work is devoted to the optimization of the glycosylation profile of a therapeutic molecule, monoclonal antibody adalimumab, at the stage of cell line cultivation, and to the study of the monoclonal antibody quality attributes that affect its biological properties. During the practical part, studies were carried out to reveal the influence of the composition of the media, feeds and additives on the glycosylation profile of the therapeutic molecule at the cultivation stage. The dependence of the biological effectiveness of the monoclonal antibody adalimumab on the composition of the glycosylated forms in the molecule was confirmed. There were also developed analytical procedures for determining the charge variants profile by cation exchange chromatography, aggregation products by size-exclusion chromatography, residual host cell DNA by real-time PCR. The procedures for determining the aggregates and residual host cell DNA were validated. The performed scientific research and the results obtained in this work can be taken into account and used as popular solutions to practical problems at the stages of technology development for the production of modern highly effective drugs.| File | Dimensione | Formato | |
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2020_10_Sterin_Tverdokhlebova.pdf
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https://hdl.handle.net/10589/166904