A HUPO TEST SAMPLE STUDY REVEALS COMMON PROBLEMS IN MASS SPECTROMETRY-BASED PROT

HUPO 测试样本研究揭示了基于质谱的 PROT 中的常见问题

• 批准号: 7957726
• 负责人: JOHN L YATES
• 金额: $ 0.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957726
• 关键词: Biology; Computer Retrieval of Information on Scientific Projects Database; Data; Databases; Environmental Pollution; Funding; Fungal Genome; Grant; Human; Institution; Ions; Laboratories; Mass Spectrum Analysis; Peptides; Proteins; Proteomics; Recombinants; Reporting; Research; Research Personnel; Resources; Sampling; Sampling Studies; Source; Testing; United States National Institutes of Health; base; improved; liquid chromatography mass spectrometry; mass spectrometer; member

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We performed a test sample study to try to identify errors leading to irreproducibility, including incompleteness of peptide sampling, in liquid chromatography-mass spectrometry-based proteomics. We distributed an equimolar test sample, comprising 20 highly purified recombinant human proteins, to 27 laboratories. Each protein contained one or more unique tryptic peptides of 1,250 Da to test for ion selection and sampling in the mass spectrometer. Of the 27 labs, members of only 7 labs initially reported all 20 proteins correctly, and members of only 1 lab reported all tryptic peptides of 1,250 Da. Centralized analysis of the raw data, however, revealed that all 20 proteins and most of the 1,250 Da peptides had been detected in all 27 labs. Our centralized analysis determined missed identifications (false negatives), environmental contamination, database matching and curation of protein identifications as sources of problems. Improved search engines and databases are needed for mass spectrometry-based proteomics.

这个子项目是众多研究子项目之一