A new approach to monitoring glycosylation during glycoprotein production

监测糖蛋白生产过程中糖基化的新方法

• 批准号: 10830193
• 负责人: Loretta Yang
• 金额: $ 98.86万
• 依托单位: GLYCOSENSORS AND DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10830193
• 关键词: Acceleration; Address; Adoption; Affect; Antibodies; Basic Science; Binding; Biological Models; Biological Products; Biological Response Modifier Therapy; Carbohydrates; Cell Culture Techniques; Chemicals; Chromatography; Complement; Consumption; Coupled; Data; Detection; Development; ERBB2 gene; Equipment; Erythropoietin; Fermentation; Flow Cytometry; Gases; Glycoproteins; Half-Life; Healthcare; Homologous Gene; Industrialization; Mammalian Cell; Mass Spectrum Analysis; Measurement; Methodology; Methods; Microspheres; Modeling; Monitor; Optics; Pattern; Performance; Pharmacologic Substance; Phase; Polysaccharides; Process; Production; Property; Protein Glycosylation; Proteins; Protocols documentation; Publishing; Quality Control; Reagent; Recombinants; Reporting; Research; Safety; Sampling; Serum; Specificity; Suspensions; Technology; Temperature; Therapeutic; Time; Trastuzumab; United States Food and Drug Administration; Universities; Variant; Work; analytical method; commercialization; cost; cost effective; design; glycosylation; immunogenicity; innovative technologies; instrument; novel; novel strategies; pharmacologic; portability; protein purification; research and development; screening; sugar; therapeutic development; tool

PROJECT SUMMARY During the production of therapeutic biologicals, glycosylation has been identified as a critical quality attribute (CQA) that must be carefully monitored to ensure that the desired protein stability, immunogenicity, antibody effector function, pharmacological safety and potency, and serum half-life are consistently obtained. Many factors can affect the composition of the glycan chains during production, such as media, temperature, dissolved gases, induction times, etc., and consequently variations in glycosylation are among the leading causes of pharmaceutical batch rejection. Currently, the characterization of protein glycosylation relies heavily on methods that employ chromatography and/or mass spectrometry, which require a high level of expertise, are time-consuming and costly, and, because they are challenging to implement during biologics production at-line, are generally performed only post-production. In recognition of the continuing need for novel analytical technologies that can be used during therapeutic production, the US Food and Drug Administration (FDA) encourages the adoption of new process analytical methods (PAT) that offer mechanisms to design, analyze, or facilitate the measurement of process parameters that affect CQAs. Here we propose to optimize and commercialize a simplified approach for monitoring glycosylation features during glycoprotein production, which we call GlycoSenseTM, that is fully aligned with the FDA PAT initiative. The GlycoSense technology uses flow cytometry with glycan-specific reagents conjugated to multiplex beads to provide a rapid, inexpensive, and easy-to-use method for monitoring protein glycosylation patterns. Flow cytometry was chosen as the preferred platform because the data are statistically robust, the equipment is relatively commonplace, the analyses consume very low quantities of sample, and it will enable Lectenz Bio to leverage its proprietary expertise in producing carbohydrate-sensing reagents. In preliminary studies reported here, GlycoSense performance metrics were established and published using glycans and glycoprotein standards. In phase II, the technology will be optimized for monitoring glycosylation changes during cell culture. This will result in GlycoSense kits that can be used for a variety of applications in basic research and in biopharma.

项目摘要 在治疗性生物制品的生产过程中，糖基化已被确定为关键质量属性 (CQA)必须仔细监测以确保所需的蛋白质稳定性、免疫原性、抗体 一致地获得效应子功能、药理学安全性和效力以及血清半衰期。许多 在生产过程中，诸如培养基，温度， 溶解气体、诱导时间等，因此糖基化的变化是 药品批次拒收的原因。 目前，蛋白质糖基化的表征严重依赖于采用色谱法的方法 和/或质谱分析，其需要高水平的专业知识，是耗时且昂贵的，并且， 由于在生物制品生产线上实施这些方法具有挑战性， 后期制作认识到对可用于生产过程的新型分析技术的持续需求， 治疗生产，美国食品和药物管理局（FDA）鼓励采用新的工艺 分析方法（PAT），提供设计、分析或促进过程测量的机制 影响CQA的参数。在这里，我们建议优化和商业化的简化方法， 在糖蛋白生产过程中监测糖基化特征，我们称之为GlycoSenseTM， 与FDA PAT计划保持一致。 GlycoSense技术使用流式细胞术，将聚糖特异性试剂与多重微珠结合 提供一种快速、廉价和易于使用的方法来监测蛋白质糖基化模式。流 选择流式细胞术作为优选的平台，因为数据在统计学上是稳健的， 相对普通，分析消耗非常少量的样品，它将使Lectenz Bio能够 利用其在生产碳水化合物传感试剂方面的专有技术。 在本文报告的初步研究中，GlycoSense性能指标是使用 聚糖和糖蛋白标准品。在第二阶段，该技术将被优化用于监测糖基化 细胞培养过程中的变化。这将导致GlycoSense试剂盒可用于多种应用， 基础研究和生物制药。