Grey Carl, Edebrink Per, Krook Maria, Jacobsson Sven P
BioPR&D, AstraZeneca R&D, SE-15185 Södertälje, Sweden.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Jul 1;877(20-21):1827-32. doi: 10.1016/j.jchromb.2009.05.003. Epub 2009 May 13.
In the present study a HPAEC-PAD method is described that was developed for monitoring the consistency of N-glycosylation during the production and purification of recombinant proteins and monoclonal antibodies. The method successfully separated 18 neutral and sialylated oligosaccharides. Results obtained were compared with MALDI-TOF MS and it was shown that both methods gave similar results. In addition, a method validation was performed showing that the HPAEC-PAD analysis was well suited for the mapping and characterization of oligosaccharides. The method was found to be robust and additionally the precision was significantly better compared to the MALDI-TOF MS method.
在本研究中，描述了一种高效阴离子交换色谱-脉冲安培检测（HPAEC-PAD）方法，该方法用于监测重组蛋白和单克隆抗体生产及纯化过程中N-糖基化的一致性。该方法成功分离了18种中性和唾液酸化的寡糖。将所得结果与基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）进行比较，结果表明两种方法给出的结果相似。此外，进行了方法验证，结果表明HPAEC-PAD分析非常适合寡糖的图谱绘制和表征。该方法被发现具有稳健性，而且与MALDI-TOF MS方法相比，精密度明显更好。
