Pande Sandhya, Rahardjo Ayu, Livingston Brittney, Mujacic Mirna
Cell Sciences and Technology, Amgen, 1201 Amgen Court W, Seattle, Washington, 98119.
Analytical Sciences, Amgen, 1201 Amgen Court W, Seattle, Washington, 98119.
Biotechnol Bioeng. 2015 Jul;112(7):1383-94. doi: 10.1002/bit.25551. Epub 2015 May 19.
Asparagine-linked glycosylation of the constant region of monoclonal antibodies (mAbs) plays an important role in their stability and efficacy and is a critical product quality attribute that needs to be consistent between various process changes and production lots. Exact product quality match is also of the utmost importance for the development of biosimilar protein therapeutics. This poses a process development challenge since mAb glycosylation profiles can fluctuate easily with changes in process parameters. Therefore, there is a need to identify methods to modulate glycosylation levels on therapeutic antibodies during a production run in order to maintain consistent product quality profiles between different drug lots. Here, we demonstrate the use of a small molecule ionophore, monensin, to increase high mannose levels on multiple therapeutic human immunoglobulins (IgGs) in both plate-based small scale production models as well as in production bioreactors. This method is simple to implement and readily applicable for multiple production cell lines. Moreover, high mannose levels can be increased without significant negative impact on titer or cell culture performance. As such, monensin gives us a manipulable product quality lever.
单克隆抗体（mAb）恒定区的天冬酰胺连接糖基化在其稳定性和效力方面发挥着重要作用，并且是一个关键的产品质量属性，需要在各种工艺变更和生产批次之间保持一致。精确的产品质量匹配对于生物类似药蛋白质疗法的开发也至关重要。这带来了工艺开发方面的挑战，因为mAb糖基化谱会随着工艺参数的变化而容易波动。因此，需要确定在生产过程中调节治疗性抗体糖基化水平的方法，以便在不同药物批次之间保持一致的产品质量谱。在此，我们展示了使用小分子离子载体莫能菌素，在基于平板的小规模生产模型以及生产生物反应器中，增加多种治疗性人免疫球蛋白（IgG）上的高甘露糖水平。该方法易于实施，并且很容易应用于多种生产细胞系。此外，可以增加高甘露糖水平，而对滴度或细胞培养性能没有显著负面影响。因此，莫能菌素为我们提供了一个可操控的产品质量杠杆。
