Bruker Daltonics Inc., San Jose, CA, USA.
Adv Exp Med Biol. 2019;1140:45-54. doi: 10.1007/978-3-030-15950-4_3.
Matrix-Assisted Laser Desorption Ionization In-Source Decay (MALDI-ISD) Mass Spectrometry is a very powerful tool for providing terminal sequence information of biomolecules with minimal sample preparations. Fragmentation is induced at the position where hydrogen radical transfers from matrix to analyte in the MALDI-ISD process by proposed mechanism. Uniform fragmentation in MALDI-ISD generates relative simple ion spectra of readable sequence ladders with labile modifications retained, which is advantageous over other fragmentation methods such as collision-induced dissociation (CID) for characterizing modifications. MALDI-ISD has been applied to de novo sequencing of a 13.6 kDa protein and fully validate sequences of therapeutic antibodies, showing its promising potential in examining reference sequences of biotherapeutics unambiguously. It has also been successfully applied to the analysis of modifications such as post-translational modifications (PTMs) and PEGylation. Here we discuss the applications of MALDI-ISD in protein sequencing and modification analysis by featuring representative studies in details.
基质辅助激光解吸电离源内裂解（MALDI-ISD）质谱是一种非常强大的工具，可在最小化样品制备的情况下提供生物分子的末端序列信息。根据提出的机制，在 MALDI-ISD 过程中，氢自由基从基质转移到分析物的位置会引发碎裂。MALDI-ISD 中的均匀碎裂会产生相对简单的可读序列梯级的离子谱，同时保留不稳定的修饰，这比其他碎裂方法（如碰撞诱导解离（CID））更有利，后者用于修饰特征化。MALDI-ISD 已应用于 13.6 kDa 蛋白质的从头测序，并完全验证了治疗性抗体的序列，这表明其在明确检查生物治疗药物的参考序列方面具有很大的潜力。它也已成功应用于修饰（如翻译后修饰（PTM）和聚乙二醇化）的分析。在这里，我们通过详细介绍代表性研究，讨论了 MALDI-ISD 在蛋白质测序和修饰分析中的应用。
