Schiel John E, Rogstad Sarah M, Boyne Michael T
National Institute of Standards and Technology, Biomolecular Measurement Division, Gaithersburg, Maryland, 20899.
Division of Pharmaceutical Research, Office of Testing and Research, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland, 20993.
J Pharm Sci. 2015 Aug;104(8):2464-72. doi: 10.1002/jps.24522. Epub 2015 Jun 5.
Monoclonal antibody therapeutics are a heterogeneous mixture of glycoforms. Multiple methods exist for defining the glycan composition and relative abundance of species present. In the current report, two MS-based methods were compared for their ability to both identify glycans and monitor differences in the glycoprofile. Gross changes in the glycoprofile can be identified either by visual inspection of fluorescence profiles and correlated to glycan identities when coupled with online MS/MS (LC-F-MS/MS) or through extracted ion chromatograms using LC-MS. In the present study, both an LC-F-MS/MS method and an automated LC-MS label free approach were able to identify minor differences in low abundance glycoforms, and data indicate a disparity in glycosylation between the analyzed batches of US and foreign-sourced mAb X. Thus, either method may be useful in characterizing monoclonal antibody therapeutics products and could serve as a potential screening test for understanding process, comparability, similarity, and possibly detecting counterfeit agents.
单克隆抗体疗法是糖型的异质混合物。存在多种用于定义存在的聚糖组成和相对丰度的方法。在本报告中，比较了两种基于质谱的方法识别聚糖和监测糖型差异的能力。糖型的总体变化可以通过荧光图谱的目视检查来识别，当与在线串联质谱（液相色谱 - 荧光 - 串联质谱，LC-F-MS/MS）结合时，可与聚糖身份相关联，或者通过使用液相色谱 - 质谱的提取离子色谱图来识别。在本研究中，液相色谱 - 荧光 - 串联质谱法和自动化的液相色谱 - 质谱无标记方法都能够识别低丰度糖型中的微小差异，数据表明分析的美国和国外来源的单克隆抗体X批次之间存在糖基化差异。因此，这两种方法都可能有助于表征单克隆抗体治疗产品，并可作为理解工艺、可比性、相似性以及可能检测假冒制剂的潜在筛选测试。
