Rapid Fc glycosylation analysis of Fc fusions with IdeS and liquid chromatography mass spectrometry
Rapid Fc glycosylation analysis of Fc fusions with IdeS and liquid chromatography mass spectrometry
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DOI：
10.4161/mabs.25302
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发表时间：
2013-09-01
期刊：
影响因子：
5.3
通讯作者：
Gong, Bing
中科院分区：
文献类型：
--
作者：
Lynaugh, Heather;Li, Huijuan;Gong, Bing
我们开发了一种快速的方法来分析Fc融合蛋白的Fc糖基化，特别是那些具有突变的Fc铰链区的融合蛋白。Fc融合蛋白用IdeS消化，IdeS是一种IgG特异性蛋白酶，具有用于底物识别和切割的外切位点。通过液相色谱质谱法直接分析所得片段。Fc聚糖的结构和相对量根据其质量和强度推导。IdeS的分离的底物识别和切割特性使得该方法适用于具有标准或非典型铰链区的广泛的Fc融合蛋白。
We developed a rapid method to analyze Fc glycosylation of Fc fusion proteins, especially those with mutated Fc hinge regions. Fc fusion proteins were digested with IdeS, an IgG specific protease with exosites for substrate recognition and cleavage. The resultant fragments were directly analyzed through liquid chromatography mass spectrometry. The structures and relative quantities of Fc glycans were deduced from their masses and intensities. The separated substrate recognition and cleavage property of IdeS makes this method applicable to a broad range of Fc fusion proteins having either standard or non-canonical hinge regions.