Molecular Structure, Amgen, Thousand Oaks, California 91320, USA.
Anal Chem. 2010 Aug 15;82(16):6751-5. doi: 10.1021/ac1013139.
Recombinant monoclonal antibodies are an important class of therapeutic agents that have found widespread use for the treatment of many human diseases. Here, we have examined the utility of ion mobility mass spectrometry (IMMS) for the rapid characterization of disulfide variants in intact IgG2 monoclonal antibodies. It is shown that IMMS reveals 2 to 3 gas-phase conformer populations for IgG2s. In contrast, a single gas-phase conformer is revealed using IMMS for both an IgG1 antibody and a Cys-232 --> Ser mutant IgG2, both of which are homogeneous with respect to disulfide bonding. This provides strong evidence that the observed IgG2 gas-phase conformers are related to disulfide bond heterogeneity. Additionally, IMMS analysis of redox enriched disulfide isoforms allows assignment of the mobility peaks to established disulfide bonding patterns. These data clearly illustrate how IMMS can be used to quickly provide information on the higher order structure of antibody therapeutics.
重组单克隆抗体是一类重要的治疗药物，已广泛用于治疗许多人类疾病。在这里，我们研究了离子淌度质谱（IMMS）在鉴定完整 IgG2 单克隆抗体中二硫键变体方面的应用。结果表明，IMMS 揭示了 IgG2 的 2 到 3 种气相构象。相比之下，使用 IMMS 仅揭示了 IgG1 抗体和 Cys-232 --> Ser 突变 IgG2 的一种气相构象，这两种抗体在二硫键连接方面都是均一的。这有力地证明了所观察到的 IgG2 气相构象与二硫键异质性有关。此外，对氧化还原富集的二硫键异构体进行 IMMS 分析，可将迁移峰分配给已建立的二硫键连接模式。这些数据清楚地说明了 IMMS 如何用于快速提供抗体治疗药物高级结构的信息。
