Model‐based optimization of antibody galactosylation in CHO cell culture

Abstract

Exerting control over the glycan moieties of antibody therapeutics is highly desirable from a product safety and batch-to-batch consistency perspective. Strategies to improve antibody productivity may compromise quality, while interventions for improving glycoform distribution can adversely affect cell growth and productivity. Process design therefore needs to consider the trade-off between preserving cellular health and productivity while enhancing antibody quality. In this work, we present a modeling platform that quantifies the impact of glycosylation precursor feedingspecifically that of galactose and uridineon cellular growth, metabolism as well as antibody productivity and glycoform distribution. The platform has been parameterized using an initial training data set yielding an accuracy of ±5% with respect to glycoform distribution. It was then used to design an optimized feeding strategy that enhances the final concentration of galactosylated antibody in the supernatant by over 90% compared with the control without compromising the integral of viable cell density or final antibody titer. This work supports the implementation of Quality by Design towards higher-performing bioprocesses. K E Y W O R D Santibody glycosylation, Chinese hamster ovary (CHO) cells, galactosylation, mathematical modeling, nucleotide sugars, process optimization

Constrained global sensitivity analysis for bioprocess design space identification

“…The set of experiments used for model construction and parameter estimation is described in [22] and The model links extracellular culture conditions, NSD metabolism in the cytosol and antibody production and glycosylation. Briefly, the cell growth model uses Monod kinetics to describe growth dependence on key metabolites.…”

Strategic Framework for Parameterization of Cell Culture Models

“…Experiments were conducted as presented in reference [28]. Briefly, fed-batch experiments with a working volume of 100 mL of cell culture were conducted in 500 mL Erlenmeyer flasks after 3 cell passages including cell revival.…”

“…A CHO cell growth, metabolism and death model previously presented in reference [28] was used for parameter estimation and sensitivity analysis. Model simulations and parameter estimation were conducted in gPROMS v.5.1.1 (Process System Enterprise Ltd., London, UK, www.psenterprise.com/ gproms).…”

“…The model examined herein describes the cell growth, metabolism and death for the CHO-T cell line. Parameter estimation has been previously performed to fit the given experimental data and examine the predictive capabilities of the model [28]. Five of the 22 replicates of galactose and uridine feeding experiments (including the control experiment) presented significant deviation from the average behaviour of the cell line.…”

Integration of Time-Series Transcriptomic Data with Genome-Scale CHO Metabolic Models for mAb Engineering

“…Chinese hamster ovary (CHO) cells are the predominant mammalian cell type used for the production of monoclonal antibody (mAb) [18][19][20]. A consensus genome-scale model, iCHO1766, was reconstructed for CHO cells in 2016 [7].…”