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Evaluation of the potential of Pap test fluid and cervical swabs to serve as clinical diagnostic biospecimens for the detection of ovarian cancer by mass spectrometry-based proteomics

机译:评估PAP试验液和宫颈拭子的潜力作为通过质谱基蛋白质组学检测卵巢癌的临床诊断生物学

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摘要

Work flow for the proteomic analysis of ovarian cancer biospecimens. a A liquid based Pap test, cervical swab (Merocel™) and snap frozen primary tumor tissue were collected from a patient with high grade serous ovarian cancer and proteins were prepared for mass spectrometry analysis as described in the Methods. b Proteins were digested with trypsin using an in-solution digest (tumor tissue extract) or FASP (Pap test fluid and swab proteins). c Peptides were HPLC fractionated and analyzed by mass spectrometry. d MS/MS data were searched against a forward and reverse sequence human Uniprot database using Sequest. Data was analyzed with Scaffold 4. Proteins were identified with at least 2 peptides at 99% probability and 0.2% decoy False Discovery Rate (FDR)
机译:卵巢癌生物组织蛋白质组学分析的工作流程。一种基于液体的PAP试验,宫颈拭子(Merocel™)和卡扣冷冻原发性肿瘤组织被从患有高级浆液卵巢癌的患者中收集,并且如方法所述,制备蛋白质进行质谱分析。使用溶液素消化B蛋白,使用溶液消化(肿瘤组织提取物)或FASP(PAP测试流体和拭子蛋白)用胰蛋白酶消化。 C肽是HPLC分级分级并通过质谱分析。使用续集,对向前和反向序列人的UniProt数据库进行搜索D MS / MS数据。用支架分析数据4.用至少2个肽在99%概率和0.2%诱饵错误发现率(FDR)中鉴定蛋白质。

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