Life Science, Upstream R&D, Merck KGaA, Darmstadt, Germany.
Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany.
Biotechnol Bioeng. 2021 May;118(5):1818-1831. doi: 10.1002/bit.27695. Epub 2021 Feb 19.
The reduction of antibody core-fucosylation is known to enhance antibody-dependent cellular cytotoxicity (ADCC). In this study, 5-Thio-l-Fucose (ThioFuc) was investigated as a media and feed supplement for modulating the fucosylation profile of therapeutic proteins and, thereby, improving the resulting effector functions. Glycan analysis of five different therapeutic proteins produced by a diverse set of Chinese hamster ovary cell lines demonstrated a clone dependent impact of ThioFuc treatment. Using rituximab as a model, an efficient dose- and time-dependent reduction of core-fucosylation up to a minimum of 5% were obtained by ThioFuc. Besides a concomitant increase in the afucosylation level up to 48%, data also revealed up to 47% incorporation of ThioFuc in place of core-fucosylation. In accordance with the glycan data, antibodies produced in the presence of ThioFuc revealed an enhanced FcγRIIIa binding up to 7.7-fold. Furthermore, modified antibodies subjected to a cell-based ADCC reporter bioassay proved to exert both a 1.5-fold enhanced ADCC efficacy and 2.6-fold enhancement in potency in comparison to their native counterparts-both of which contribute to an improvement in the ADCC activity. In conclusion, ThioFuc is a potent fucose derivative with potential applications in drug development processes.
抗体核心岩藻糖基化的减少已知可增强抗体依赖性细胞毒性 (ADCC)。在这项研究中，5-硫代-l-岩藻糖 (ThioFuc) 被用作调节治疗性蛋白岩藻糖基化谱的培养基和饲料补充剂，从而改善产生的效应功能。对五种不同的治疗性蛋白进行的聚糖分析，这些蛋白由不同的中国仓鼠卵巢细胞系产生，表明 ThioFuc 处理对克隆有依赖性影响。以利妥昔单抗为模型，通过 ThioFuc 获得了高达 5%的有效剂量和时间依赖性核心岩藻糖基化降低，同时伴随高达 48%的去岩藻糖基化水平增加，数据还显示高达 47%的 ThioFuc 取代核心岩藻糖基化。与聚糖数据一致，在 ThioFuc 存在下产生的抗体显示出 FcγRIIIa 结合增强高达 7.7 倍。此外，经过细胞 ADCC 报告生物测定的修饰抗体被证明与天然抗体相比具有 1.5 倍增强的 ADCC 功效和 2.6 倍增强的效力，这两者都有助于提高 ADCC 活性。总之，ThioFuc 是一种有效的岩藻糖衍生物，具有在药物开发过程中的潜在应用。
