Department of Biomedical Engineering, Rutgers The State University of New Jersey, 599 Taylor Road, Piscataway, NJ 08854, USA.
Department of Chemical & Biochemical Engineering, Rutgers The State University of New Jersey, 98 Brett Road, Piscataway, New Jersey 08854, USA.
J Pharm Sci. 2023 Jun;112(6):1485-1491. doi: 10.1016/j.xphs.2023.01.012. Epub 2023 Jan 20.
N-glycosylation is a post-translational modification that occurs during the production of monoclonal antibody (mAb) therapeutics. During production of mAb based therapeutics the use of various hosts and cell culture additives attribute to glycan heterogeneity. The safety and efficacy of monoclonal antibodies with mechanism of actions that utilize Fc effector functions can be negatively impacted by glycan heterogeneity and thus is often considered a critical quality attribute (CQA). In this mini review, we discuss recent advances in mAb sample preparation specifically focused on denaturation, enzymatic processing, and released glycans derivatization methods. Additionally, we review the recent advances in characterization of released and intact N-glycans using chromatography, capillary electrophoresis, and mass spectrometry techniques with a focus on rapid, automated approaches that support analysis of glycosylation profiles of biopharmaceuticals. We delve into advances within sample preparation techniques that allow for rapid and robust sample preparation as well as how these techniques are being used for innovative at-line high-throughput screening and process analytical technology (PAT). The future of biomanufacturing is focused on decreasing process costs while increasing process understanding and quality for novel biologic candidates and biosimilars. Therefore, advances in PAT for biotherapeutics will positively influence current manufacturing practices and enable further bioprocess automation.
N-糖基化是一种翻译后修饰，发生在单克隆抗体 (mAb) 治疗药物的生产过程中。在 mAb 治疗药物的生产过程中，各种宿主和细胞培养添加剂的使用导致聚糖异质性。作用机制利用 Fc 效应子功能的单克隆抗体的安全性和疗效可能会受到聚糖异质性的负面影响，因此通常被认为是关键质量属性 (CQA)。在这篇综述中，我们讨论了 mAb 样品制备的最新进展，特别是针对变性、酶处理和释放糖基化衍生化方法的进展。此外，我们还综述了使用色谱法、毛细管电泳法和质谱法对释放和完整 N-糖进行表征的最新进展，重点介绍了支持生物制药糖基化谱分析的快速、自动化方法。我们深入探讨了样品制备技术的进展，这些技术允许快速、稳健的样品制备，以及这些技术如何用于创新的在线高通量筛选和过程分析技术 (PAT)。生物制造的未来重点是降低工艺成本，同时提高新型生物候选物和生物类似物的工艺理解和质量。因此，生物治疗剂的 PAT 进展将积极影响当前的制造实践，并实现进一步的生物过程自动化。
