The extent and pattern of glycosylation on therapeutic antibodies can influence their circulatory half-life, engagement of effector functions, and immunogenicity, with direct consequences to efficacy and patient safety. Hence, controlling glycosylation patterns is central to any drug development program, yet poses a formidable challenge to the bio-manufacturing industry. Process changes, which can affect glycosylation patterns, range from manufacturing at different scales or sites, to switching production process mode, all the way to using alternative host cell lines. In the emerging space of biosimilars development, often times all of these aspects apply. Gaining a deep understanding of the direction and extent to which glycosylation quality attributes can be modulated is key for efficient fine-tuning of glycan profiles in a stage appropriate manner, but establishment of such platform knowledge is time consuming and resource intensive. Here we report an inexpensive and highly adaptable screening system for comprehensive modulation of glycans on antibodies expressed in CHO cells. We characterize 10 media additives in univariable studies and in combination, using a design of experiments approach to map the design space for tuning glycosylation profile attributes. We introduce a robust workflow that does not require automation, yet enables rapid process optimization. We demonstrate scalability across deep wells, shake flasks, AMBR-15 cell culture system, and 2 L single-use bioreactors. Further, we show that it is broadly applicable to different molecules and host cell lineages. This universal approach permits fine-tuned modulation of glycan product quality, reduces development costs, and enables agile implementation of process changes throughout the product lifecycle.

治疗性抗体上糖基化的程度和方式可影响其循环半衰期，效应器功能的参与和免疫原性，直接影响疗效和患者安全性。因此，控制糖基化模式对于任何药物开发计划都是至关重要的，但对生物制造行业提出了巨大的挑战。可能影响糖基化模式的过程变化，从不同规模或位置的制造，一直到转换生产过程模式，一直到使用替代宿主细胞系。在生物仿制药发展的新兴空间中，通常所有这些方面都适用。深入了解糖基化质量属性可以被调节的方向和程度，对于以适当阶段的方式有效地微调聚糖谱至关重要，但是建立此类平台知识既耗时又耗费资源。在这里，我们报告了一种廉价且高度适应的筛选系统，可对CHO细胞中表达的抗体上的聚糖进行全面调节。我们使用一种实验设计方法来映射设计空间以调整糖基化谱图属性，从而在单变量研究和组合分析中表征10种培养基添加剂。我们引入了强大的工作流程，该流程不需要自动化，但可以实现快速的流程优化。我们展示了深孔，摇瓶，AMBR-15细胞培养系统和2 L一次性生物反应器的可扩展性。进一步，我们表明它广泛适用于不同的分子和宿主细胞谱系。这种通用方法允许对聚糖产品质量进行微调，降低开发成本，并在整个产品生命周期中灵活地实施过程变更。