Glycan analysis of glycoprotein pharmaceuticals: Evaluation of analytical approaches to Z number determination in pharmaceutical erythropoietin products |
| |
Authors: | Chun-Ting Yuen Yong Zhou Qing-Zhou Wang Ji-Feng Hou Adrian Bristow Jun-Zhi Wang |
| |
Institution: | aNational Institute for Biological Standards and Control (NIBSC), TDI/Protein Science, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK;bNational Institute for Control of Pharmaceutical and Biological Products (NICPBP), No.2, Tiantan Xili, Chongwen District, Beijing 100050, PR China |
| |
Abstract: | N-Glycosylation of many glycoprotein drugs is important for biological activity and should therefore be the target of specific and quantitative analytical methods. In this study, we focus on the two N-glycan mapping approaches that are used in pharmacopoeial monograph to analyse N-glycans released from fifteen preparations of recombinant human erythropoietin supplied by ten Chinese manufacturers. Underivatised N-glycans were analysed by high performance anion-exchange chromatography with pulsed amperometric detection and fluorophore-labelled N-glycans were analysed by weak anion-exchange and normal-phase high performance liquid chromatography. N-glycans were also analysed by matrix assisted laser desorption ionisation mass spectrometry. The release of N-glycans by PNGase F was shown to be consistent. Z number, a mathematical expression of the total negatively charged N-glycans composition has provided a convenient way to summarise the complex dataset and it might be suitable for product consistency monitoring. However, this Z number reduces the information of individual acidic N-glycan structure and is also found to be method dependent. Therefore, its use requires clear specification and validation. In this study, we only found weak but positive correlation between the Z number and its bioactivity. Wide range of N-glycans yields were obtained from the fifteen preparations but the significance of their differences is unclear. |
| |
Keywords: | N-glycans analysis Glycotherapeutics EPO Recombinant erythropoietin Z number |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|