糖基化是目前生物制药开发中非常感兴趣的话题,因为它与药物安全性和有效性有关。这项工作描述了NISTmAb初级样品(PS)糖基化的实验室间研究的结果,单克隆抗体参考材料。来自工业界的76个实验室,大学,研究,政府,和欧洲的医院部门,北美,亚洲,澳大利亚共提交了103份关于聚糖分布的报告。本研究的主要目的是报告和比较目前用于mAb糖基化分析的全部分析方法的结果。因此,参与不受限制,实验室选择自己的测量技术。以各种方式确定蛋白质糖基化,包括完整的单克隆抗体水平,蛋白质片段,糖肽,或释放的聚糖,使用各种各样的衍生方法,分离,identification,和量化。因此,结果的多样性是巨大的,每个实验室鉴定的聚糖组成数量为4至48。总的来说,报告了116种聚糖成分,其中57种组合物可以被指定为共有丰度值。这些共识中位数为NISTmAbPS提供了社区衍生的价值。与共识中位数的协议不取决于具体方法或实验室类型。该研究提供了生物糖基化测量的最新技术,并明确需要协调糖基化分析方法。
Glycosylation is a topic of intense current interest in the development of biopharmaceuticals because it is related to drug safety and efficacy. This work describes results of an interlaboratory
study on the glycosylation of the Primary Sample (PS) of NISTmAb, a monoclonal antibody reference material. Seventy-six laboratories from industry, university, research, government, and hospital sectors in Europe, North America, Asia, and Australia submitted a total of 103 reports on
glycan distributions. The principal objective of this
study was to report and compare results for the full range of analytical methods presently used in the glycosylation analysis of mAbs. Therefore, participation was unrestricted, with laboratories choosing their own measurement techniques. Protein glycosylation was determined in various ways, including at the level of intact mAb, protein fragments, glycopeptides, or released glycans, using a wide variety of methods for derivatization, separation, identification, and quantification. Consequently, the diversity of results was enormous, with the number of
glycan compositions identified by each laboratory ranging from 4 to 48. In total, one hundred sixteen glycan compositions were reported, of which 57 compositions could be assigned consensus abundance values. These consensus medians provide community-derived values for NISTmAb PS. Agreement with the consensus medians did not depend on the specific method or laboratory type. The
study provides a view of the current state-of-the-art for biologic glycosylation measurement and suggests a clear need for harmonization of glycosylation analysis methods.