Gangwar Neelesh, Dixit Neha, Rathore Anurag S
School of Interdisciplinary Research, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India.
Department of Chemical Engineering, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India.
Appl Microbiol Biotechnol. 2025 Jan 22;109(1):16. doi: 10.1007/s00253-025-13405-5.
Monoclonal antibodies are extensively used as biotherapeutics for treatment of a variety of diseases. Glycosylation of therapeutic antibodies is considered a critical quality attribute as it influences the effector function, circulatory half-life, immunogenicity, and eventually efficacy and patient safety. During upstream process development, media components play a significant role in determining the glycosylation profile. In this study, we have evaluated 20 media additives (metal ions, vitamins, sugars, nucleosides). Six of the additives were shortlisted for their impact and then used to modulate the glycosylation profile of an in-house produced mAb (G0 2.38 ± 0.08%, G0F 75.58 ± 0.45%, G1F 10.07 ± 0.04%, G2F 0.54 ± 0.01%, G0F-N 5.84 ± 0.32%, sialylation 1.60 ± 0.33%, mannosylation 1.56 ± 0.39%) to achieve the glycan profile of a commercially available reference product (G0 2.49 ± 0.07%, G0F 37.83 ± 0.37%, G1F 34.77 ± 0.03%, G2F 4.87 ± 0.01%, G0F-N 2.34 ± 0.12%, sialylation 9.84 ± 0.30%, mannosylation 2.86 ± 0.29%). The proposed approach yielded us a glycan profile (G0 2.10 ± 0.07%, G0F 38.00 ± 0.49%, G1F 31.92 ± 0.09%, G2F 5.26 ± 0.54%, G0F-N 1.92 ± 0.02%, sialylation 10.28 ± 1.68%, mannosylation 3.12 ± 0.29%) that was near identical to that of the reference product. Equally importantly, other quality attributes including charge variants, aggregates, titer, and viability were not found to be significantly impacted by the addition of the additives under consideration. KEY POINTS: • Screened 20 media additives to evaluate their effect on glycosylation of mAbs. • Developed glycosylation indices models to evaluate the effect of various additives. • Additive concentrations were optimized to target the reference product profile.
单克隆抗体被广泛用作治疗多种疾病的生物疗法。治疗性抗体的糖基化被认为是一个关键的质量属性，因为它会影响效应功能、循环半衰期、免疫原性，并最终影响疗效和患者安全。在上游工艺开发过程中，培养基成分在决定糖基化谱方面起着重要作用。在本研究中，我们评估了20种培养基添加剂（金属离子、维生素、糖类、核苷）。其中六种添加剂因其影响而入围，然后用于调节内部生产的单克隆抗体的糖基化谱（G0 2.38±0.08%，G0F 75.58±0.45%，G1F 10.07±0.04%，G2F 0.54±0.01%，G0F-N 5.84±0.32%，唾液酸化1.60±0.33%，甘露糖基化1.56±0.39%），以实现市售参考产品的聚糖谱（G0 2.49±0.07%，G0F 37.83±0.37%，G1F 34.77±0.03%，G2F 4.87±0.01%，G0F-N 2.34±0.12%，唾液酸化9.84±0.30%，甘露糖基化2.86±0.29%）。所提出的方法为我们提供了一个与参考产品几乎相同的聚糖谱（G0 2.10±0.07%，G0F 38.00±0.49%，G1F 31.92±0.09%，G2F 5.26±0.54%，G0F-N 1.92±0.02%，唾液酸化10.28±1.68%，甘露糖基化3.12±0.29%）。同样重要的是，未发现所考虑的添加剂的添加对包括电荷变体、聚集体、滴度和活力在内的其他质量属性有显著影响。要点如下：•筛选了20种培养基添加剂以评估它们对单克隆抗体糖基化的影响。•开发了糖基化指数模型以评估各种添加剂的效果。•优化添加剂浓度以靶向参考产品谱。
