原句结构不变：Serial multi-omic analysis of proteome, phosphoproteome, and acetylome provides insights into changes in protein expression, cell signaling, cross-talk and epigenetic pathways involved in disease pathology and treatment. However, ubiquitylome and HLA peptidome data collection used to understand protein degradation and antigen presentation have not together been serialized, and instead require separate samples for parallel processing using distinct protocols. Here we present MONTE, a highly sensitive multi-omic native tissue enrichment workflow, that enables serial, deep-scale analysis of HLA-I and HLA-II immunopeptidome, ubiquitylome, proteome, phosphoproteome, and acetylome from the same tissue sample. We demonstrate that the depth of coverage and quantitative precision of each 'ome is not compromised by serialization, and the addition of HLA immunopeptidomics enables the identification of peptides derived from cancer/testis antigens and patient specific neoantigens. We evaluate the technical feasibility of the MONTE workflow using a small cohort of patient lung adenocarcinoma tumors.© 2023. The Author(s). 蛋白质组、磷酸化蛋白组和乙酰化蛋白组的串联多组学分析可研究疾病病理学和治疗中与蛋白质表达、细胞信号传递、跨谈和表观遗传途径有关的变化。但是，为了理解蛋白质降解和抗原呈递，泛素组和HLA肽组数据采集仍需使用不同的协议并分开取样进行平行加工，未能进行串联。在此，我们提供了MONTE，一种高灵敏度的多组学天然组织富集工作流程，可以从同一组织样本中对HLA-I和HLA-II免疫肽组、泛素组、蛋白质组、磷酸化蛋白组和乙酰化蛋白组进行串联、深度规模的分析。我们证明了每个“组学”的覆盖深度和定量精确度不会因为串联而受损，HLA免疫肽组学的添加可以识别出来源于癌症/睾丸抗原和患者特异性新抗原的肽段。我们使用小型肺腺癌患者队列评估了MONTE工作流程的技术可行性。© 2023. 作者。

Serial multi-omic analysis of proteome, phosphoproteome, and acetylome provides insights into changes in protein expression, cell signaling, cross-talk and epigenetic pathways involved in disease pathology and treatment. However, ubiquitylome and HLA peptidome data collection used to understand protein degradation and antigen presentation have not together been serialized, and instead require separate samples for parallel processing using distinct protocols. Here we present MONTE, a highly sensitive multi-omic native tissue enrichment workflow, that enables serial, deep-scale analysis of HLA-I and HLA-II immunopeptidome, ubiquitylome, proteome, phosphoproteome, and acetylome from the same tissue sample. We demonstrate that the depth of coverage and quantitative precision of each 'ome is not compromised by serialization, and the addition of HLA immunopeptidomics enables the identification of peptides derived from cancer/testis antigens and patient specific neoantigens. We evaluate the technical feasibility of the MONTE workflow using a small cohort of patient lung adenocarcinoma tumors.© 2023. The Author(s).