Complementary MS methods assist conformational characterization of antibodies with altered S-S bonding networks.
Complementary MS methods assist conformational characterization of antibodies with altered S-S bonding networks.
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DOI：
10.1007/s13361-013-0582-4
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发表时间：
2013-06
影响因子：
3.2
通讯作者：
Gross, Michael L.
中科院分区：
文献类型：
--
作者：
Jones, Lisa M.;Zhang, Hao;Cui, Weidong;Kumar, Sandeep;Sperry, Justin B.;Carroll, James A.;Gross, Michael L.
关键词：
随着治疗性单克隆抗体（mAb）成为生物技术的主要焦点和下一代药物的来源，需要新的分析方法或组合方法来监测高阶结构的变化和翻译后修饰的影响。这些分子的复杂性及其对结构变化的脆弱性提供了一个严峻的挑战。我们在此描述了互补质谱法的使用，该方法不仅表征突变mAb，而且还可以为表征其他蛋白质治疗剂和生物仿制药的高阶结构提供一般框架。为了构建挑战，我们选择了具有不同二硫键异构体结构的IgG2亚类成员作为模型，以评估使用离子迁移率、自上而下的MS测序和蛋白质快速光化学氧化（FPOP）形式的蛋白质足迹的整体方法。这三种方法快速、灵敏，可响应IgG 2的Cys→Ser突变体构象的细微变化，每种突变体代表一种单一的二硫键同种型，并且可串联使用以探测更高级结构。该结果表明，这种结合使用各种方法的方法可以帮助蛋白质治疗剂的开发和质量控制。
As therapeutic monoclonal antibodies (mAbs) become a major focus in biotechnology and a source of the next-generation drugs, new analytical methods or combination methods are needed for monitoring changes in higher order structure and effects of post-translational modifications. The complexity of these molecules and their vulnerability to structural change provide a serious challenge. We describe here the use of complementary mass spectrometry methods that not only characterize mutant mAbs but also may provide a general framework for characterizing higher order structure of other protein therapeutics and biosimilars. To frame the challenge, we selected members of the IgG2 subclass that have distinct disulfide isomeric structures as a model to evaluate an overall approach that uses ion mobility, top-down MS sequencing, and protein footprinting in the form of fast photochemical oxidation of proteins (FPOP). These three methods are rapid, sensitive, respond to subtle changes in conformation of Cys→Ser mutants of an IgG2, each representing a single disulfide isoform, and may be used in series to probe higher order structure. The outcome suggests that this approach of using various methods in combination can assist the development and quality control of protein therapeutics.
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影响因子：
120.7
作者：
Giezen, Thijs J.;Mantel-Teeuwisse, Aukje K.;Egberts, Antoine C. G.
通讯作者：
Egberts, Antoine C. G.
影响因子：
46.9
作者：
Aggarwal, Saurabh
通讯作者：
Aggarwal, Saurabh
影响因子：
7.4
作者：
Gau, Brian C.;Chen, Hao;Zhang, Yun;Gross, Michael L.
通讯作者：
Gross, Michael L.
影响因子：
8
作者：
Baerga-Ortiz, A;Hughes, CA;Komives, EA
通讯作者：
Komives, EA
影响因子：
7.4
作者：
Jones, Lisa M.;Sperry, Justin B.;Carroll, James A.;Gross, Michael L.
通讯作者：
Gross, Michael L.