Cheon, Dong Huey Nam, Eun Ji Park, Kyu Hyung Woo, Se Joon Lee, Hye Jin Kim, Hee Cheol Yang, Eun Gyeong Lee, Cheolju Lee, Ji Eun 2024-01-20T05:04:09Z 2024-01-20T05:04:09Z 2021-09-03 2016-01 1535-3893 https://pubs.kist.re.kr/handle/201004/124546 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. English AMER CHEMICAL SOC HAPTOGLOBIN-RELATED PROTEIN AMINO-ACID-SEQUENCE HUMAN SERUM PROTEOME OVARIAN-CANCER INTACT PROTEIN BREAST-CANCER QUANTITATIVE-ANALYSIS BIOMARKER DISCOVERY LUNG-CANCER TRANSTHYRETIN Comprehensive Analysis of Low-Molecular-Weight Human Plasma Proteome Using Top-Down Mass Spectrometry Article 10.1021/acs.jproteome.5b00773 1 JOURNAL OF PROTEOME RESEARCH, v.15, no.1, pp.229 - 244 JOURNAL OF PROTEOME RESEARCH 15 1 229 244 scie scopus 000367706000018 2-s2.0-84953238347 Biochemical Research Methods Biochemistry & Molecular Biology Article HAPTOGLOBIN-RELATED PROTEIN AMINO-ACID-SEQUENCE HUMAN SERUM PROTEOME OVARIAN-CANCER INTACT PROTEIN BREAST-CANCER QUANTITATIVE-ANALYSIS BIOMARKER DISCOVERY LUNG-CANCER TRANSTHYRETIN 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)