Fast analysis of recombinant monoclonal antibodies using IdeS proteolytic digestion and electrospray mass spectrometry
Fast analysis of recombinant monoclonal antibodies using IdeS proteolytic digestion and electrospray mass spectrometry
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DOI：
10.1016/j.ab.2011.04.030
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发表时间：
2011-08-15
影响因子：
2.9
通讯作者：
Bihoreau, Nicolas
中科院分区：
文献类型：
--
作者：
Chevreux, Guillaume;Tilly, Nolwenn;Bihoreau, Nicolas
我们描述了一种快速和信息丰富的方法来研究单克隆抗体（MAbs）的翻译后修饰。MAb首先被特异性酶消化，该酶切割铰链结构域下的重链。二硫键还原后，释放出约25 kDa的三条多肽链，并通过液相色谱-质谱法（LC-MS）进行分析。通过在MS分析前平分重链，该方法提供了更好的MS分辨率，并有助于研究N-连接聚糖以及其他修饰（C-末端赖氨酸缺失）。焦谷氨酰胺化和氧化）。样品制备和分析可以在几个小时内完成。(C)2011 Elsevier Inc. All rights reserved.
We describe a fast and informative method to investigate the posttranslational modifications of monoclonal antibodies (MAbs). The MAb is first digested by a specific enzyme that cleaves heavy chains under the hinge domain. After reduction of disulfide bridges, three polypeptide chains of approximately 25 kDa are released and analyzed by liquid chromatography-mass spectrometry (LC-MS). By bisecting the heavy chains prior to MS analysis, this method provides a better MS resolution and facilitates the study of the N-linked glycans as well as of other modifications (loss of C-terminal lysine. pyroglutamination, and oxidation). The sample preparation and analysis can be performed within few hours. (C) 2011 Elsevier Inc. All rights reserved.