Advances of Glycometabolism Engineering in Chinese Hamster Ovary Cells

Abstract

As the most widely used mammalian cell line, Chinese hamster ovary (CHO) cells can express various recombinant proteins with a post translational modification pattern similar to that of the proteins from human cells. During industrial production, cells need large amounts of ATP to support growth and protein expression, and since glycometabolism is the main source of ATP for cells, protein production partly depends on the efficiency of glycometabolism. And efficient glycometabolism allows less glucose uptake by cells, reducing production costs, and providing a better mammalian production platform for recombinant protein expression. In the present study, a series of progresses on the comprehensive optimization in CHO cells by glycometabolism strategy were reviewed, including carbohydrate intake, pyruvate metabolism and mitochondrial metabolism. We analyzed the effects of gene regulation in the upstream and downstream of the glucose metabolism pathway on cell’s growth and protein expression. And we also pointed out the latest metabolic studies that are potentially applicable on CHO cells. In the end, we elaborated the application of metabolic models in the study of CHO cell metabolism.

Developing microRNAs as engineering tools to modulate monoclonal antibody galactosylation

“…Therefore, engineering the amount of expressed galactosylated glycans on the mAb provides the possibility to actively modulate their therapeutic effects in patients and to ensure consistent and controllable product quality. Most approaches have focused mainly on overexpression or silencing of key genes in relevant metabolic pathways or variation of feeding‐strategies and media supplementation (Amann et al, 2018; Bydlinski et al, 2018; Chung, Wang, Yang, Ponce, et al, 2017; Gramer et al, 2011; Prabhu et al, 2018, 2022; Zhang et al, 2021). However, current genetic engineering approaches based on knock‐out and overexpression are often unable to fine‐tune the phenotype of glycosylation.…”

Advancements in CHO metabolomics: techniques, current state and evolving methodologies

“…These sugar precursors can be majorly detected using GC-MS except few (UDP-Gal, UDP-galactose, UDP-glucose) which were uniquely identified in studies performed using GC-MS, whereas NMR was unable to detect these polar metabolites ( Supplementary Table S1 ). An efficient glycometabolism for protein production allows less glucose uptake by cells ( Zhang et al, 2021 ). Besides, reduction of lactate levels in CHO cell cultures improves the product titre by 11%–32% without having significant impact on cell growth suggesting that reduced level of lactate in culture potentially either due to its consumption or inhibited generation may be the primary cause of its beneficial effects ( Ahn and Antoniewicz, 2011 ; Naik et al, 2023 ).…”

“…The glycolytic pathway and its associated pathways upregulates in the exponential phase of culture to potentially meet the increased demand of energy during cell proliferation (Sengupta et al, 2011;Templeton et al, 2013;Zhu et al, 2022;Naik et al, 2023) (Table 3; Figure 3). For example, glucose, glucose-6-phosphate, pyruvate, phosphoenolpyruvate, fructose 1,6-bisphosphate, and fructose 6phosphate increases in the exponentially growing cells (Buchsteiner et al, 2018;Zhang et al, 2021;Coulet et al, 2022). Majority of these metabolites can be detected using GC-MS, except glucose which can be universally identified using any of the three techniques (LC-MS, GC-MS, NMR) (Supplementary Table S1).…”

CircRNA-0013747 induces mesangial cell proliferation in IgA nephropathy by targeting the Warburg effect via miR-330- 3p/PKM2 signaling

“…However, when cells are under hypoxic conditions, pyruvic acid is not converted into acetyl-coenzyme A but rather into lactic acid, a process known as anaerobic cellular respiration [10]. In this scenario, the net energy balance is reduced to just two ATP molecules, making anaerobic glucose metabolism energetically ine cient [11]. In the early 1920s, Warburg, Posener, and Negelein made noteworthy discoveries regarding the behavior of tumor tissue in vitro.…”