NAD+ supplementation improves mAb productivity in CHO cells via a glucose metabolic shift

Abstract

Aerobic glycolysis and its by-product lactate accumulation are usually associated with adverse culture phenotypes such as poor cell viability and productivity. Due to the lack of knowledge on underlying mechanisms and accompanying biological processes, the regulation of aerobic glycolysis has been an ongoing challenge in culture process development for therapeutic protein productivity. Nicotinamide adenine dinucleotide (NAD⁺ ), a coenzyme and co-substrate in energy metabolism, promotes the conversion of inefficient glycolysis into an efficient oxidative phosphorylation (OXPHOS) pathway. However, the effect of NAD⁺ on Chinese hamster ovary (CHO) cells for biopharmaceutical production has not been reported yet. In this work, we aimed to elucidate the influence of NAD⁺ on cell culture performance by examining metabolic shifts and mAb productivity. The supplementation of NAD⁺ increased the intracellular concentration of NAD⁺ and promoted SIRT3 expression. Antibody titer and the specific productivity in the growth phase were improved by up to 1.82- and 1.88-fold, respectively, with marginal restrictions on cell growth. NAD⁺ significantly reduced the accumulation of reactive oxygen species (ROS) and the lactate yield from glucose, determined by lactate accumulation versus glucose consumption (Y_LAC/GLC ). In contrast, OXPHOS capacity and amino acid consumption rate increased substantially. Collectively, these results suggest that NAD⁺ contributes to improving therapeutic protein productivity in bioprocessing via inducing an energy metabolic shift.

Keywords: Chinese hamster ovary cells; SIRT3; aerobic glycolysis; metabolic shift; nicotinamide adenine dinucleotide (NAD); therapeutic antibody productivity.