Current Approaches to Glycoprotein Analysis
Current Approaches to Glycoprotein Analysis
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DOI：
10.2174/0929866521666140626095752
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发表时间：
2014-01-01
影响因子：
1.6
通讯作者：
Liu, Li
中科院分区：
文献类型：
--
作者：
Hagan, Andrew K.;Wang, Maomao;Liu, Li
糖蛋白在治疗中变得越来越相关，包括用于治疗心肌梗塞和中风的组织纤溶酶原激活剂、用于贫血的促红细胞生成素和用于癌症的各种基于单克隆抗体的治疗。蛋白质N-和O-糖基化可能是蛋白质经历的最关键和极其复杂的翻译后修饰，其表征提出了一个重大挑战。本文将讨论当前用于糖蛋白表征的技术，重点关注可用的治疗性糖蛋白。关键的分析技术，如高效液相色谱（HPLC），毛细管电泳（CE），质谱（MS）将被描述，以及必要的化学标记方法的灵敏检测。将讨论从蛋白质中释放寡糖的成熟的化学和酶促方法，以及基于非特异性蛋白酶彻底水解蛋白质的更现代的方法。
Glycoproteins are becoming increasingly relevant in therapeutics, including tissue plasminogen activator for the treatment of myocardial infarction and strokes, erythropoietin for anemia and various monoclonal antibody-based treatments for cancer. Protein N- and O-glycosylation is perhaps the most crucial and immensely complex posttranslational modification that proteins undergo, and its characterization presents a major challenge. This review will discuss current techniques for the characterization of glycoproteins, with a focus on therapeutic glycoproteins where available. The crucial analytical techniques, such as high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), and mass spectrometry (MS) will be described, alongside the necessary chemical labeling methods for sensitive detection. The well-established chemical and enzymatic methods for oligosaccharide release from proteins will be discussed, as will more modern methods based on exhaustive protein hydrolysis with non-specific proteases.