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Abstract：
Amyloidosis is a disease characterized by local and systemic extracellular deposition of amyloid protein fibrils where its excessive accumulation in tissues and resistance to degradation can lead to organ failure. Diagnosis is challenging because of approximately 36 different amyloid protein subtypes. Imaging methods like immunohistochemistry and the use of Congo red staining of amyloid proteins for laser capture microdissection combined with liquid chromatography tandem mass spectrometry (LMD/LC-MS/MS) are two diagnostic methods currently used depending on the expertise of the pathology laboratory. Here, we demonstrate a streamlined in situ amyloid peptide spatial mapping by Matrix Assisted Laser Desorption Ionization-Mass Spectrometry Imaging (MALDI-MSI) combined with Trapped Ion Mobility Spectrometry for potential transthyretin (ATTR) amyloidosis subtyping. While we utilized the standard LMD/LC-MS/MS workflow for amyloid subtyping of 31 specimens from different organs, we also evaluated the potential introduction in the MS workflow variations in data acquisition parameters like dynamic exclusion, or testing Data Dependent Acquisition combined with High-Field Asymmetric Waveform Ion Mobility Spectrometry (DDA FAIMS) versus Data Independent Acquisition (DIA) for enhanced amyloid protein identification at shorter acquisition times. We also demonstrate the use of Mascot\'s Error Tolerant Search and PEAKS de novo sequencing for the sequence variant analysis of amyloidosis specimens.
摘要：
淀粉样变是一种以淀粉样蛋白原纤维的局部和全身细胞外沉积为特征的疾病,其中淀粉样蛋白原纤维在组织中的过度积累和对降解的抗性可导致器官衰竭。由于大约36种不同的淀粉样蛋白亚型，诊断具有挑战性。成像方法，如免疫组织化学和使用刚果红染色淀粉样蛋白进行激光捕获显微切割结合液相色谱串联质谱（LMD/LC-MS/MS）是目前使用的两种诊断方法，具体取决于病理学实验室的专业知识。这里,我们通过基质辅助激光解吸电离-质谱成像（MALDI-MSI）结合陷阱离子迁移谱技术对潜在的转甲状腺素蛋白（ATTR）淀粉样变性亚型进行了简化的原位淀粉样肽空间定位。虽然我们利用标准LMD/LC-MS/MS工作流程对来自不同器官的31个样本进行淀粉样蛋白亚型分型，我们还评估了MS工作流程中数据采集参数变化的潜在引入，如动态排除，或测试数据相关采集结合高场非对称波形离子迁移谱（DDAFAIMS）与数据独立采集（DIA）相结合，以在更短的采集时间内增强淀粉样蛋白识别。我们还证明了Mascot的容错搜索和PEAKS从头测序用于淀粉样变性标本的序列变异分析。
