Institute of Technology and Science, The University of Tokushima, Minamijosanjima-cho 2-1, Tokushima 770-8506, Japan.
Advanced Technology and Science, The University of Tokushima, Minamijosanjima-cho 2-1, Tokushima 770-8506, Japan.
J Biosci Bioeng. 2014 May;117(5):632-8. doi: 10.1016/j.jbiosc.2013.10.022. Epub 2013 Dec 4.
The aggregation of therapeutic antibodies during the manufacturing process is problematic because of the potential risks posed by the aggregates, such as an unexpected immune response. One of the hallmark effects of trehalose, a disaccharide consisting of two alpha-glucose units, is as a chemical chaperone with anti-aggregation activity. In this study, Chinese hamster ovary (CHO) cell line producing a diabody-type bispecific antibody were cultured in medium containing trehalose and the aggregation of the secreted proteins during the culture process was analyzed. An analysis of the various forms of the antibody (monomeric, dimeric, and large aggregates) showed that trehalose decreased the relative content of large aggregates by two thirds. The aggregation kinetics indicated that trehalose directly inhibited the polymerization and aggregation steps in a nucleation-dependent aggregation mechanism. Moreover, both specific and volumetric antibody production were increased in CHO cells cultured in trehalose-containing medium. Thus, the addition of trehalose to recombinant CHO cell cultures would offer a practical strategy for quality improvement in the production of therapeutic antibodies.
在制造过程中，治疗性抗体的聚集是一个问题，因为聚集物可能带来潜在风险，例如意外的免疫反应。海藻糖是一种由两个α-葡萄糖单元组成的二糖，其标志之一是具有抗聚集活性的化学伴侣。在这项研究中，使用含有海藻糖的培养基培养产生二价抗体的中国仓鼠卵巢（CHO）细胞系，并分析培养过程中分泌蛋白的聚集情况。对抗体的各种形式（单体、二聚体和大聚集体）的分析表明，海藻糖使大聚集体的相对含量减少了三分之二。聚集动力学表明，海藻糖在依赖成核的聚集机制中直接抑制聚合和聚集步骤。此外，在含有海藻糖的培养基中培养的 CHO 细胞中，特异性和比容抗体的产生均增加。因此，向重组 CHO 细胞培养物中添加海藻糖将为提高治疗性抗体生产的质量提供一种实用策略。
