采用 IdeS 酶和液相色谱-质谱联用技术快速分析 Fc 融合蛋白的 Fc 糖基化。
Rapid Fc glycosylation analysis of Fc fusions with IdeS and liquid chromatography mass spectrometry.
机构信息
GlycoFi; Biologics Discovery; Merck & Co. Inc.; Lebanon, NH USA.
出版信息
MAbs. 2013 Sep-Oct;5(5):641-5. doi: 10.4161/mabs.25302. Epub 2013 Jun 20.
Abstract
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.
摘要
我们开发了一种快速分析 Fc 融合蛋白 Fc 糖基化的方法，特别是那些 Fc 铰链区发生突变的蛋白。用 IdeS（一种 IgG 特异性蛋白酶，具有底物识别和切割的外切位点）将 Fc 融合蛋白进行酶切。通过液相色谱-质谱直接分析酶切片段。根据分子量和强度推断出 Fc 聚糖的结构和相对含量。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。
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