Barnett Institute of Chemical and Biological Analysis, Northeastern University, Boston, Massachusetts 02115, United States.
Anal Chem. 2012 Sep 18;84(18):7638-42. doi: 10.1021/ac301840f. Epub 2012 Aug 29.
Capillary electrophoresis with laser induced fluorescence detection (CE-LIF) was employed for rapid sialic acid speciation, facilitating the quantitative determination of N-glycolylneuraminic acid (Neu5Gc) and N-acetylneuraminic acid (Neu5Ac) on glycoproteins. Derivatization of the sialic acids with 2-aminoacridone (2-AMAC), using classical reductive amination in a nonaqueous solvent, led to the spontaneous decarboxylation of the sialic acid residues as determined by CE-LIF and offline mass spectrometric analysis. Modification of both the labeling conditions, to drive the decarboxylation reaction to completion and the CE-LIF parameters to separate the neutral species by complexation with a neutral coated capillary and borate reversed polarity, led to a robust platform for the rapid, sensitive, and quantitative speciation of sialic acids. The method can readily be used for quality control of recombinant biopharmaceuticals.
毛细管电泳激光诱导荧光检测(CE-LIF)被用于快速唾液酸特异性分析,有助于定量测定糖蛋白中的 N-羟乙酰神经氨酸(Neu5Gc)和 N-乙酰神经氨酸(Neu5Ac)。唾液酸与 2-氨基吖啶酮(2-AMAC)衍生化,在非水溶剂中通过经典的还原胺化作用,导致唾液酸残基自发脱羧,通过 CE-LIF 和离线质谱分析确定。修饰标记条件,使脱羧反应完全,以及 CE-LIF 参数通过与中性涂层毛细管和硼酸反转极性形成复合物来分离中性物质,为快速、灵敏和定量的唾液酸特异性分析提供了一个强大的平台。该方法可用于重组生物制药的质量控制。