Targeted proteomics and miRNomics to tackle biomarkers of infectious and neurodegenerative diseases and cancer

目前，分子生物标志物的研究已被广泛认为是改善感染性、神经退行性疾病和癌症的预防、诊断、预后、监测和治疗的基础。本项目重点开发生物体液中蛋白质、糖蛋白和miRNA生物标志物的纯化、预富集、分离、鉴定和表征的创新分析方法，特别是与COVID-19 （spike糖蛋白和核衣壳蛋白）、败血症（原降钙素蛋白）、帕金森病和阿尔茨海默病（α -突触核蛋白和簇蛋白糖蛋白）相关的蛋白质、糖蛋白和miRNA生物标志物。慢性淋巴细胞白血病（miR21）和结直肠癌（癌胚抗原糖蛋白，miR-421， miR130b-3p和miR27a-3p）。不同的策略将被研究，包括在完整水平上直接分析生物标志物，或通过自下而上的方法，在酶消化后获得特征片段（如肽、磷酸肽、糖肽或聚糖）。同样，潜在生物标志物的结构、组成或浓度的可能差异和改变将通过对照和病理样本的比较来评估。为此，它将主要使用高分辨率微尺度分离技术耦合质谱（毛细管液相色谱和毛细管电泳耦合质谱，CapLC-MS和CE-MS）的全扫描MS和串联MS/MS模式，以及在线纯化和预富集技术，如在线固相萃取毛细管电泳-质谱（在线SPE-CE-MS）。最后，先进的数据分析工具的应用将为不同的基于ms的比较研究中产生的大量复杂多维数据提供更系统和全面的解释。

Nowadays, research on molecular biomarkers is widely recognized as fundamental to improving the prevention, diagnosis, prognosis, monitoring and treatment of infectious and neurodegenerative diseases, and cancer. This project is focused on the development of innovative analytical methods for purification, preconcentration, separation, identification and characterization in biological fluids of proteins, glycoproteins and miRNA biomarkers, specifically related to COVID-19 (spike glycoprotein and nucleocapside protein), sepsis (procalcitonin protein), Parkinsons and Alzheimers diseases (alpha-synuclein protein and clusterin glycoprotein, respectively), chronic lymphocytic leukemia (miR21) and colorectal cancer (carcinoembryonic antigen glycoprotein, miR-421, miR130b-3p and miR27a-3p). Different strategies will be investigated, including the direct analysis of the biomarkers at the intact level or by bottom-up approaches, after their enzymatic digestion to obtain characteristic fragments (e.g. peptides, phosphopeptides, glycopeptides or glycans). Likewise, possible differences and alterations in the structure, composition or concentration of the potential biomarkers will be evaluated from a comparison of control and pathological samples. For these purposes, it will be primarily used high-resolution microscale separation techniques coupled to mass spectrometry (capillary liquid chromatography and capillary electrophoresis coupled to mass spectrometry, CapLC-MS and CE-MS) in full scan MS and tandem MS/MS modes, and online purification and preconcentration techniques, such as on-line solid-phase extraction capillary electrophoresis-mass spectrometry (on-line SPE-CE-MS). Finally, the application of advanced data analysis tools will provide a more systematic and comprehensive interpretation of the enormous amount of complex multidimensional data generated in the different comparative MS-based studies.