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MALDI-target integrated platform for affinity-captured protein digestion

机译:用于亲和力捕获的蛋白质消化的MALDI-target集成平台

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摘要

To address immunocapture of proteins in large cohorts of clinical samples high throughput sample processing is required. Here a method using the proteomic sample platform, 1SET (integrated selective enrichment target) that integrates highly specific immunoaffinity capture of protein biomarker, digestion and sample cleanup with a direct interface to mass spectrometry is presented. The robustness of the on-ISET protein digestion protocol was validated by MALDI MS analysis of model proteins, ranging from 40 fmol to 1 pmol per nanovial. On-ISET digestion and MALDI MS/MS analysis of immunoaffinity captured disease-associated biomarker PSA (prostate specific antigen) from human seminal plasma are presented.
机译:为了解决大批临床样品中蛋白质的免疫捕获问题,需要高通量样品处理。本文介绍了一种使用蛋白质组学样品平台1SET(集成的选择性富集靶标)的方法,该方法将蛋白质生物标志物的高特异性免疫亲和捕获,消化和样品净化与质谱的直接接口相结合。通过MALDI MS分析模型蛋白验证了on-ISET蛋白消化方案的鲁棒性,范围为每纳小瓶40 fmol至1 pmol。介绍了人精浆中免疫亲和力捕获的疾病相关生物标志物PSA(前列腺特异性抗原)的ISET消化和MALDI MS / MS分析。

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