期刊
JOURNAL OF PROTEOME RESEARCH
卷 15, 期 1, 页码 229-244出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.5b00773
关键词
human plasma; low-molecular-weight (LMW) plasma proteins; cleaved products; top-down mass spectrometry; gel-eluted liquid fraction entrapment electrophoresis (GELPrEE); post-translational modification (PTM); single amino acid variation (SAAV); biomarker; colorectal cancer (CRC)
资金
- Multiomics program through NRF - Korean MSIP [NRF-2012M3A9B9036679]
- KIST institutional program [2E25495]
While human plasma serves as a great source for disease diagnosis, low-molecular-weight (LMVV) proteome (<30 kDa) has been shown to contain a rich source of diagnostic biomarkers. Here we employ top-down mass spectrometry to analyze the LMW proteoforms present in four types of human plasma samples pooled from three healthy controls (HCs) without immunoaffinity depletion and with depletion of the top two, six, and seven high-abundance proteins. The LMVV proteoforms were first fractionated based on molecular weight using gel-eluted liquid fraction entrapment electrophoresis (GELFrEE). Then, the GELFrEE fractions containing up to 30 kDa were subjected to nanocapillary-LC-MS/MS, and the high-resolution MS and MS/MS data were processed using ProSightPC 3.0. As a result, a total of 442 LMW proteins and cleaved products, including those with post-translational modifications and single amino acid variations, were identified. From additional comparative analysis of plasma samples without immunoaffinity depletion between HCs and colorectal cancer (CRC) patients via top-down approach, tens of LMW proteoforms, including platelet factor 4, were found to show >1.5-fold changes between the plasma samples of HCs and CRC patients, and six of the LMVV proteins were verified by Western blot analysis.
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