Along with mass spectrometry (MS), ion mobility separations (IMS) are advancing to ever larger biomolecules. The emergence of electrospray ionization (ESI) and native MS enabled the IMS/MS analyses of proteins up to ∼100 kDa in the 1990s and whole protein complexes and viruses up to ∼10 MDa since the 2000s. Differential IMS (FAIMS) is substantially orthogonal to linear IMS based on absolute mobility K and offers exceptional resolution, unique selectivity, and steady filtering readily compatible with slower analytical methods such as electron capture or transfer dissociation (ECD/ETD). However, the associated MS stages had limited FAIMS to ions with m/z < 8000 and masses under ∼300 kDa. Here, we integrate high-definition FAIMS with the Q-Exactive Orbitrap UHMR mass spectrometer that can handle m/z up to 80,000 and MDa-size ions in the native ESI regime. In the initial evaluation, the oligomers of monoclonal antibody adalimumab (148 kDa) are size-selected up to at least the nonamers (1.34 MDa) with m/z values up to ∼17,000. This demonstrates the survival and efficient separation of noncovalent MDa assemblies in the FAIMS process, opening the door to novel analyses of the heaviest macromolecules.

中文翻译：

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将差分离子淌度分离扩展到兆道尔顿范围

与质谱 (MS) 一起，离子淌度分离 (IMS) 正在向更大的生物分子迈进。电喷雾电离 (ESI) 和天然 MS 的出现使得 IMS/MS 分析的蛋白质在 20 世纪 90 年代高达 ∼100 kDa，自 2000 年代以来能够对整个蛋白质复合物和病毒进行高达 ∼10 MDa 的分析。差分 IMS (FAIMS) 与基于绝对迁移率K的线性 IMS 基本正交，并提供卓越的分辨率、独特的选择性和稳定的过滤，易于与电子捕获或转移解离 (ECD/ETD) 等较慢的分析方法兼容。然而，相关的 MS 阶段将 FAIMS 限制为m / z < 8000 和质量低于 ∼300 kDa 的离子。在这里，我们将高清 FAIMS 与 Q-Exactive Orbitrap UHMR 质谱仪集成在一起，该质谱仪可以在原生 ESI 范围内处理高达 80,000 个m / z和 MDa 大小的离子。在初步评估中，单克隆抗体阿达木单抗寡聚物 (148 kDa) 的大小选择至少为九聚物 (1.34 MDa)，m / z值高达 ~17,000。这证明了 FAIMS 过程中非共价 MDa 组装体的存活和有效分离，为最重大分子的新颖分析打开了大门。