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<p>Adenosine-5’-triphosphate (ATP) serves as biological energy used in various cellular reactions, and maintaining the concentration of ATP is essential for cellular homeostasis. It has been estimated that the synthesis of a single antibody molecule requires approximately 5,000 ATP molecules. Therefore, increasing the intracellular ATP level in recombinant protein-expressing cells can be a promising strategy to enhance productivity. This study aims to verify the effects of four purine precursors—adenine, adenosine, hypoxanthine, and inosine—on culture performance through metabolite and productivity analyses. Supplementation of purine precursors significantly increased intracellular ATP levels as well as specific productivity by up to 5.6-fold in a dose-dependent manner. The effects of precursor supplementation varied among cell lines, with greater improvements observed in the low-productivity cell line B. In batch cultures, adenine supplementation increased the monoclonal antibody (mAb) titer by 27.8%, while in pseudo-perfusion cultures, it led to a 51.6% increase. Specific mAb productivity was enhanced by 2.92-fold and 1.61-fold under these conditions, respectively. A positive correlation was observed between specific mAb productivity and intracellular ATP levels, as well as with C-source usage capacity, suggesting that precursor supplementation may enhance cellular carbon metabolism, including glycolysis and mitochondrial oxidative phosphorylation. In conclusion, these results indicate the potential application of purine precursors as media additives or feed components for enhancing productivity in bioproduction processes.</p>

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Purine precursor supplementation increases productivity of antibody-expressing Chinese Hamster Ovary cells by elevating intracellular ATP level

  • 期刊论文

  • 发表日期：

    2026年7月6日

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  • Jong Uk Park,
  • Joo Hee Bae,
  • Hyun Gyu Yu,
  • Hye-Jin Han,
  • Hagyeong Kim,
  • Jong Youn Baik

摘要

Adenosine-5’-triphosphate (ATP) serves as biological energy used in various cellular reactions, and maintaining the concentration of ATP is essential for cellular homeostasis. It has been estimated that the synthesis of a single antibody molecule requires approximately 5,000 ATP molecules. Therefore, increasing the intracellular ATP level in recombinant protein-expressing cells can be a promising strategy to enhance productivity. This study aims to verify the effects of four purine precursors—adenine, adenosine, hypoxanthine, and inosine—on culture performance through metabolite and productivity analyses. Supplementation of purine precursors significantly increased intracellular ATP levels as well as specific productivity by up to 5.6-fold in a dose-dependent manner. The effects of precursor supplementation varied among cell lines, with greater improvements observed in the low-productivity cell line B. In batch cultures, adenine supplementation increased the monoclonal antibody (mAb) titer by 27.8%, while in pseudo-perfusion cultures, it led to a 51.6% increase. Specific mAb productivity was enhanced by 2.92-fold and 1.61-fold under these conditions, respectively. A positive correlation was observed between specific mAb productivity and intracellular ATP levels, as well as with C-source usage capacity, suggesting that precursor supplementation may enhance cellular carbon metabolism, including glycolysis and mitochondrial oxidative phosphorylation. In conclusion, these results indicate the potential application of purine precursors as media additives or feed components for enhancing productivity in bioproduction processes.