Seong, Yunseo Yoo, Young Sook Akter, Hafeza Kang, Min-Jung 2024-01-20T01:02:05Z 2024-01-20T01:02:05Z 2021-09-04 2017-08 1570-0232 https://pubs.kist.re.kr/handle/201004/122496 Proteomics is a valuable approach to discover biomarkers in human plasma for early diagnosis. However, detection of biomarkers in the plasma is still challenging because of its large protein content. In our study, we established albumin/IgG depletion methods for identification of low abundance proteins using two commercial kits with additional buffer conditions and various concentrations of cold acetone. Trypsin digestion, desalting, and data-dependent acquisition were also optimized. More than 80% depletion of albumin/IgG was achieved with two commercial kits and 98% depletion of albumin was obtained with 70% cold acetone. Recovery of four reference proteins, BNP (47-76), insulin, cytochrome c, and ubiquitin was obtained in all optimized methods. The best recovery of reference proteins was obtained using the ProteoExtract albumin/IgG removal kit with buffer A (61%-106%). After cold acetone precipitation, three reference proteins were recovered more than 48% except ubiquitin (12%). The number of identified proteins by Mascot was 28, 35, 17, and 34 for ProteoExtract, ProteoPrep, 70%, and 50% cold acetone, respectively. Furthermore, optimized methods detected MS/MS fragmentation patterns of elevated BNP in patient samples with cardiac disease. Our study provides the conditions for efficient biomarker discovery by minimal removing of high abundant proteins. English Elsevier BV Sample preparation for detection of low abundance proteins in human plasma using ultra-high performance liquid chromatography coupled with highly accurate mass spectrometry Article 10.1016/j.jchromb.2017.06.023 1 Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, v.1060, pp.272 - 280 Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 1060 272 280 N scie scopus 000411540100036 2-s2.0-85021088184 Biochemical Research Methods Chemistry, Analytical Biochemistry & Molecular Biology Chemistry Article ACUTE MYOCARDIAL-INFARCTION CANCER BIOMARKER DISCOVERY PROSTATE-SPECIFIC ANTIGEN HUMAN SERUM GEL-ELECTROPHORESIS PROTEOMIC ANALYSIS TROPONIN ASSAYS TASK-FORCE DEPLETION UPDATE Biomarkers Human plasma Low abundance proteins LTQ orbitrap Sample preparation