S‐adenosylmethionine and methylthioadenosine boost cellular productivities of antibody forming Chinese hamster ovary cells

Abstract

The improvement of cell specific productivities for the formation of therapeutic proteins is an important step towards intensified production processes. Among others, the induction of the desired production phenotype via proper media additives is a feasible solution provided that said compounds adequately trigger metabolic and regulatory programs inside the cells. In this study, S-(5′-adenosyl)-L-methionine (SAM) and 5′-deoxy-5′-(methylthio)adenosine (MTA) were found to stimulate cell specific productivities up to approx. 50% while keeping viable cell densities transiently high and partially arresting the cell cycle in an anti-IL-8-producing CHO-DP12 cell line. Noteworthy, MTA turned out to be the chemical degradation product of the methyl group donor SAM and is consumed by the cells.

Methylthioadenosine (MTA) boosts cell‐specific productivities of Chinese hamster ovary cultures: dosage effects on proliferation, cell cycle and gene expression

“…All sampling and measurement procedures were performed with three technical replicates. The determination of cell cycle distribution was performed as described before [21]. All sampling and measurement procedures were performed with two technical replicates.…”

“…Own studies have already revealed that MTA addition diminished growth, increased CSP, altered cell cycle phases and cell size [21]. Consequently, MTA should be considered as a multi-layer regulator of cell growth, cell cycle, and protein formation that is a highly promising additive for boosting CSP.…”

“…Single MTA addition to the medium increases CSP in CHO cells. Furthermore, cells demonstrated cell cycle arrest and increased cell size [21]. Growth arrest induction is a common strategy to increase CSP [22].…”

Compartment-specific metabolome labeling enables the identification of subcellular fluxes that may serve as promising metabolic engineering targets in CHO cells

“…To date, the compartment-specific analytical approach has shown its suitability for multiple metabolomic studies investigating CHO cells under in vivo-like conditions [8,15,[24][25][26][27][28][29][30]. The latter is enabled by fast and standardized metabolism inactivation.…”

Compartment‐specific 13C metabolic flux analysis reveals boosted NADPH availability coinciding with increased cell‐specific productivity for IgG1 producing CHO cells after MTA treatment

“…However, the underlying mechanisms that explain the phenotype remain fragmented. Recently, Verhagen et al (2020a,b) [28,29] demonstrated that cell-cycle arrest could be achieved by exposing CHO cells to the effector 5′-deoxy-5′-(methylthio)adenosine (MTA), which increased CSP by 50%.…”