Since production of therapeutics with the help of living organisms is the core of biopharmaceuticals, a lot of attention must be paid to regulating the quality of the obtained product. One of the most important quality attributes of recombinant monoclonal antibodies (mAb) is fucosylation, which strongly influences the cell-mediated cytotoxicity response (ADCC) in patients. The degree of mAb fucosylation can be regulated during the upstream production process, by varying the culture conditions, and thereby directing cellular metabolism towards the desired product quality. The goal of the doctoral thesis was to deepen the understanding of cellular metabolism of CHO culture during fed-batch to support modulation of mAb fucosylation by the means of bioinformatics analysis of metabolomics data. Metabolomic data were analyzed with a mixture of univariate and multivariate statistics and specialized bioinformation approaches, that enabled identification of four different metabolic phases during fed-batch, as well as the list of metabolites that were differed significantly between the standard bioprocess and the one, which was optimized to obtain a higher degree of fucosylated mAbs. We have thus improved the understanding of metabolic processes that take place within the CHO cells during the fed-batch bioprocess and found conditions that allow the production of mAbs with varying degree of fucosylation.
|
