Abstract：
Proteomic analysis of formalin-fixed paraffin-embedded (FFPE) tumor tissue specimens has gained interest in the last 5 years due to technological advances and improved sample collection, as well as biobanking for clinical trials. The real-world implementation of clinical proteomics to these specimens, however, is hampered by tedious sample preparation steps and long instrument acquisition times.
To advance the translation of quantitative proteomics into the clinic, we are comparing the performance of the leading commercial nanoflow liquid chromatography (nLC) system (based on literature reviews), the Easy-nLC 1200 (Thermo Fisher Scientific, Waltham, MA, U.S.A.), to the Evosep One HPLC (Evosep Biosystems, Odense, Denmark). We measured FFPE-tissue digests from 21 biological replicates with a similar gradient on both of the LC systems while keeping the on-column amount (1 µg total protein) and the single-shot data-dependent acquisition-based MS/MS method constant.
Overall, the Evosep One facilitates robust and sensitive high-throughput sample acquisition, making it suitable for clinical MS. We found the Evosep One to be a useful platform for positioning mass spectrometry-based proteomics in the clinical setting. The clinical application of nLC/MS will inform clinical decision-making in oncology and other diseases.
To advance the translation of quantitative proteomics into the clinic, we are comparing the performance of the leading commercial nanoflow liquid chromatography (nLC) system (based on literature reviews), the Easy-nLC 1200 (Thermo Fisher Scientific, Waltham, MA, U.S.A.), to the Evosep One HPLC (Evosep Biosystems, Odense, Denmark). We measured FFPE-tissue digests from 21 biological replicates with a similar gradient on both of the LC systems while keeping the on-column amount (1 µg total protein) and the single-shot data-dependent acquisition-based MS/MS method constant.
Overall, the Evosep One facilitates robust and sensitive high-throughput sample acquisition, making it suitable for clinical MS. We found the Evosep One to be a useful platform for positioning mass spectrometry-based proteomics in the clinical setting. The clinical application of nLC/MS will inform clinical decision-making in oncology and other diseases.
摘要：
■由于技术进步和样品收集的改进，福尔马林固定石蜡包埋（FFPE）肿瘤组织标本的蛋白质组学分析在过去5年中引起了人们的兴趣，以及临床试验的生物库。临床蛋白质组学对这些标本的实际应用，然而,繁琐的样品制备步骤和长的仪器采集时间阻碍。
■为了推进定量蛋白质组学在临床中的应用，我们正在比较领先的商用纳米低液相色谱（nLC）系统的性能（基于文献综述），Easy-nLC1200(ThermoFisherScientific，沃尔瑟姆,MA,U.S.A.),到EvosepOneHPLC(EvosepBiosystems，欧登塞,丹麦)。我们在两个LC系统上以相似的梯度测量了21个生物重复的FFPE组织消化物，同时保持柱上量（1µg总蛋白）和基于单次数据依赖性采集的MS/MS方法恒定。
■总的来说，EvosepOne有助于稳健和灵敏的高通量样品采集，使其适合临床MS。我们发现EvosepOne是在临床环境中定位基于质谱的蛋白质组学的有用平台。nLC/MS的临床应用将为肿瘤学和其他疾病的临床决策提供信息。
■为了推进定量蛋白质组学在临床中的应用，我们正在比较领先的商用纳米低液相色谱（nLC）系统的性能（基于文献综述），Easy-nLC1200(ThermoFisherScientific，沃尔瑟姆,MA,U.S.A.),到EvosepOneHPLC(EvosepBiosystems，欧登塞,丹麦)。我们在两个LC系统上以相似的梯度测量了21个生物重复的FFPE组织消化物，同时保持柱上量（1µg总蛋白）和基于单次数据依赖性采集的MS/MS方法恒定。
■总的来说，EvosepOne有助于稳健和灵敏的高通量样品采集，使其适合临床MS。我们发现EvosepOne是在临床环境中定位基于质谱的蛋白质组学的有用平台。nLC/MS的临床应用将为肿瘤学和其他疾病的临床决策提供信息。
