Chemmalil Letha, Kulkarni Tanmay, Raman Mathura, Singh Priya, Qian Yueming, Chumsae Chris, McHugh Kyle, Huang Zhuangrong, Hodgman Eric, Borys Michael C, Ding Julia, Li Gloria, Leone Anthony
Biological Process Analytical Group Bristol Myers Squibb Devens Massachusetts USA.
Analytical Science & Technology Bristol Myers Squibb Devens Massachusetts USA.
Eng Life Sci. 2025 Jan 23;25(1):e202400031. doi: 10.1002/elsc.202400031. eCollection 2025 Jan.
This study emphasizes the critical importance of closely monitoring and controlling the sialic acid content in therapeutic glycoproteins, including EPO, interferon-γ, Orencia, Enbrel, and others, as the level of sialylation directly impacts their pharmacokinetics (PK), immunogenicity, potency, and overall clinical performance due to its influence on protein clearance via hepatic asialoglycoprotein receptors (ASGPR). The ASGPR recognizes and binds to glycoproteins exposed to terminal galactose or N-acetylgalactosamine residues, leading to receptor-mediated endocytosis. Recent studies have demonstrated that sialylation of O-linked glycan plays a role in protecting against macrophage galactose lectin (MGL)-mediated clearance. In addition to the impact on serum half-life, sialylation can influence other clinical outcomes, including immunogenicity, potency, and cytotoxicity. Therefore, the level of sialic acid is a critical quality attribute (CQA), and monitoring and regulating sialylation has become a regulatory requirement to ensure desired clinical performance. To achieve consistent levels of sialic acid-to-protein ratio, the time of upstream harvest and conductivity of downstream wash buffers must be tightly regulated based on the sialic acid content. Therefore, the utilization of process analytical technology (PAT) tools for generating real-time or near-real-time sialic acid content is a business-critical requirement. This work demonstrates the utility of an integrated PAT system for near real-time online sialic acid measurements. The system consists of a micro-sequential injection analyzer (µSIA) interfaced with SegFlow and an ultra performance liquid chromatography (UPLC). The fully automated architecture exemplifies the execution of online sampling, automatic sample preparation, and subsequent online UPLC analysis. This carefully orchestrated PAT framework effectively supports the requirements of QbD-driven continuous bioprocessing.
本研究强调了密切监测和控制治疗性糖蛋白（包括促红细胞生成素、干扰素-γ、阿巴西普、恩利等）中唾液酸含量的至关重要性，因为唾液酸化水平直接影响其药代动力学（PK）、免疫原性、效力及整体临床性能，这是由于其通过肝脏去唾液酸糖蛋白受体（ASGPR）对蛋白质清除产生影响。ASGPR识别并结合暴露于末端半乳糖或N-乙酰半乳糖胺残基的糖蛋白，导致受体介导的内吞作用。最近的研究表明，O-连接聚糖的唾液酸化在防止巨噬细胞半乳糖凝集素（MGL）介导的清除中发挥作用。除了对血清半衰期的影响外，唾液酸化还可影响其他临床结果，包括免疫原性、效力和细胞毒性。因此，唾液酸水平是一个关键质量属性（CQA），监测和调节唾液酸化已成为确保期望临床性能的一项监管要求。为了实现唾液酸与蛋白质比例的一致水平，必须根据唾液酸含量严格控制上游收获时间和下游洗涤缓冲液的电导率。因此，利用过程分析技术（PAT）工具来生成实时或近实时的唾液酸含量是一项对业务至关重要的要求。这项工作展示了一个用于近实时在线唾液酸测量的集成PAT系统的实用性。该系统由与SegFlow接口的微顺序注射分析仪（µSIA）和超高效液相色谱（UPLC）组成。全自动架构体现了在线采样、自动样品制备及后续在线UPLC分析的执行过程。这个精心编排的PAT框架有效地支持了质量源于设计（QbD）驱动的连续生物加工的要求。
