Impact of Copper and Melezitose Supplementation on Adalimumab Production and N-linked Glycosylation in Chinese Hamster Ovary Cell Culture

Information from the abstract

Controlling mAb N-linked glycosylation is essential for ensuring therapeutic safety and efficacy. Because culture supplements can differentially affect both glycosylation profiles and culture performance, empirical optimization is often necessary. In this study, fed-batch shake-flask cultures of a CHO cell clone producing adalimumab, a biosimilar of the anti-TNF-α antibody Humira, were supplemented with copper or melezitose at specific concentrations. The goal was to increase G1F and decrease hybrid (e.g., -NAF) glycan levels to better match the innovator profile. Effects on cell growth and productivity were also investigated. Copper supplementation at 1.5 mM slightly increased G1F glycans, decreased hybrid glycans (e.g., NAF), and increased Man5 glycans. Similarly, 25 mM melezitose also slightly increased G1F glycans, although it reduced Man5 levels and had little to no effect on hybrid glycans. Melezitose slightly decreased cell growth and productivity, whereas copper enhanced productivity. These findings are directly applicable to clinical mAb candidates. Although only single concentrations of copper and melezitose were tested, these results suggest that copper and melezitose supplementation can be used as levers to modulate glycosylation.

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Related topics: Viral Infectious Diseases and Gene Expression in Insects · Monoclonal and Polyclonal Antibodies Research · Protein purification and stability