Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland.
Biotechnol Bioeng. 2018 Jun;115(6):1378-1393. doi: 10.1002/bit.26567. Epub 2018 Mar 31.
As a key parameter impacting functional and structural heterogeneity, protein glycosylation is a critical quality attribute for antibody biotherapeutic manufacturing. The glycan patterns on recombinant antibodies, particularly on the conserved fragment crystallizable (Fc) region, can have significant effects on an antibody's functional activities including clearance rate, antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and anti-inflammatory activity. In this review, we examined specific glycan attachments (fucosylation, sialylation, galactosylation, high-mannose, and bisecting glycans) and their importance to antibody properties. Next, we summarized the recent and current achievements on controlling antibody glycoforms in Chinese hamster ovary (CHO) and other mammalian cells through multiple strategies including genetic engineering, protein engineering, media modification, and other emerging technologies. Further, the impact of one carbohydrate modification on other glycan structures is also described. Finally, approaches to generate desirable homogenous glycan profiles on antibodies are also detailed. By applying multiple complementary intracellular and extracellular strategies, biotechnologists are well on their ways to precisely tuning antibody glycoforms emerging from bioreactors in the coming decades.
作为影响功能和结构异质性的关键参数，蛋白质糖基化是抗体生物治疗制造的关键质量属性。重组抗体上的聚糖模式，特别是在保守的片段结晶（Fc）区域，对抗体的功能活性有显著影响，包括清除率、抗体依赖性细胞毒性（ADCC）、补体依赖性细胞毒性（CDC）和抗炎活性。在这篇综述中，我们研究了特定的聚糖附着（岩藻糖基化、唾液酸化、半乳糖基化、高甘露糖和双分支聚糖）及其对抗体性质的重要性。接下来，我们总结了通过多种策略，包括遗传工程、蛋白质工程、培养基修饰和其他新兴技术，在 CHO 和其他哺乳动物细胞中控制抗体糖型的最新和当前进展。此外，还描述了一种碳水化合物修饰对其他聚糖结构的影响。最后，还详细介绍了生成抗体上理想均一糖型谱的方法。通过应用多种互补的细胞内和细胞外策略，生物技术人员在未来几十年内，将很好地实现从生物反应器中精确调整抗体糖型的目标。
