Monoclonal antibodies (MAbs), a class of commercially valuable biologics with drug sales in the US exceeding $24.6 billion in 2012[1], are used for treating a wide range of diseases such as cancer, psoriasis and Crohn’s disease. A vast majority of MAbs are expressed in mammalian cell lines such as Chinese Hamster Ovary (CHO) cells that generate human-like protein structures and enable proper post-translational modifications. One such post-translational modification that results in structural and pharmacological changes in the protein is glycosylation, involving the addition of a sugar moiety (glycan) to the protein backbone. Of particular interest is N-linked glycosylation, a post-translational modification involving the addition of the glycan to an Asn-X-Ser/Thr sequon in the MAb. The non-template driven enzymatic attachment of different glycans to the MAb yields a heterogeneous glycosylation profile which can alter the product quality of the MAb, compromising the efficacy and safety of the drug product, and hence the product quality. Therefore to maintain product quality and a consistent glycoform profile it is necessary to identify the process factors that can affect glycosylation. Previous studies have shown that nutrient supplements added to cell culture media can affect the glycosylation profile in MAbs. In this work, we present a rational approach to control the final glycosylation profile in MAbs using appropriate amino acid supplementation. First we evaluate the effect of 11 different amino acids on the glycan distribution in MAbs using a Box-Behnken experimental design. Subsequently, we carry out a controllability analysis as introduced in St. Amand et al [2] in order to identify which glycans can be controlled by supplementing the media with specific amino acids. Finally, we design and evaluate via simulation, a controller to direct the glycoform profile in MAb to a desired state by manipulating the appropriate amino acid concentrations identified by the controllability analysis. 1. Aggarwal S (2014) What's fueling the biotech engine-2012 to 2013. Nat Biotech 32: 32-39. 2. St. Amand MM, Tran K, Radhakrishnan D, Robinson AS, Ogunnaike BA (2014) Controllability Analysis of Protein Glycosylation in Cho Cells. PLoS ONE 9: e87973.

Controlling the glycosylation profile in MAbs by amino acid supplementation

249th National Meeting (2015)

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