Glycoprofiling effects of media additives on IgG produced by CHO cells in fed‐batch bioreactors

Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

Section: Discussionmentioning

confidence: 99%

Section: Galactosylationmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

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Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

“…As the activity of a mAb drug is commonly mediated through antibody-dependent cellular cytotoxicity (ADCC) or complementdependent cytotoxicity, the antibody glycoform can affect these processes by either facilitating or hindering recruitment of necessary interactors. 14,15 Aglycosylation, where the protein lacks a glycan residue at the Asn 297 site, is another possible outcome. 11 The glycosylation state of the protein can additionally affect stability, immunogenicity, and clearance rate.…”

Section: Introductionmentioning

confidence: 99%

Multivariate data analysis of growth medium trends affecting antibody glycosylation

Powers

Trunfio

Velugula‐Yellela

et al. 2019

Use of multivariate data analysis for the manufacturing of biologics has been increasing due to more widespread use of data-generating process analytical technologies (PAT) promoted by the US FDA. To generate a large dataset on which to apply these principles, we used an in-house model CHO DG44 cell line cultured in automated micro bioreactors alongside PAT with four commercial growth media focusing on antibody quality through N-glycosylation profiles. Using univariate analyses, we deter-

“…Our research addressed the need to better understand how complex input variables within the biomanufacturing process affect product quality. CQAs are physical, chemical, biological, or microbiological properties that must be within specified ranges to ensure the desired product quality, and in the case of mAb products one particularly important CQA is the distribution of product glycoforms . N ‐linked glycosylation may affect various therapeutic properties of the antibody, from effector function, immunogenicity, stability, and clearance rate .…”

Section: Introductionmentioning

confidence: 99%

Real‐time quantification and supplementation of bioreactor amino acids to prolong culture time and maintain antibody product quality

Powers

Wang

Fratz‐Berilla

et al. 2019

Real‐time monitoring of cell cultures in bioreactors can enable expedited responses necessary to correct potential batch failure perturbations which may normally go undiscovered until the completion of the batch and result in failure. Currently, analytical technologies are dedicated to real‐time monitoring of bioreactor parameters such as pH, dissolved oxygen, and temperature, nutrients such as glucose and glutamine, or metabolites such as lactate. Despite the importance of amino acids as the building blocks of therapeutic protein products, other than glutamine their concentrations are not commonly measured. Here, we present a study into amino acid monitoring, supplementation strategies, and how these techniques may impact the cell growth profiles and product quality. We used preliminary bioreactor runs to establish baselines by determining initial amino acid consumption patterns, the results of which were used to select a pool of amino acids which gets depleted in the bioreactor. These amino acids were combined into blends which were supplemented into bioreactors during a subsequent run, the concentrations of which were monitored using a mass spectrometry based at‐line method we developed to quickly assess amino acid concentrations from crude bioreactor media. We found that these blends could prolong culture life, reversing a viable cell density decrease that was leading to batch death. Additionally, we assessed how these strategies might impact protein product quality, such as the glycan profile. The amino acid consumption data were aligned with the final glycan profiles in principal component analysis to identify which amino acids are most closely associated with glycan outcomes.