title: Developing a strong anion exchange/RP (SAX/RP) 2D LC system for high-abundance proteins depletion in human plasma.
abstract: Human plasma is dominated by high-abundance proteins which severely impede the detection of low-abundance proteins. Unfortunately, now there is no efficient method for large-scale depletion of high-abundance proteins in human plasma. In this study, we developed a new strategy, strong anion exchange (SAX)/RP 2D LC system, which has potential for large-scale depletion of high-abundance proteins in human plasma. Separation gradients of the system were optimized to ensure an extensive separation of plasma proteins. Plasma was fractionated into 67 fractions by SAX. All these fractions were subjected a thorough separation by the 2D RPLC and 66 peaks with high UV absorption (>20 mAU) at 215 nm were collected. Proteins in these peaks were identified by LC-MS/MS analysis. Results showed that 83 proteins could be identified in these peaks, 68 among them were reported to be high- or middle-abundance proteins in plasma. All these proteins had definite retention times and were mapped in the 2D SAX-RP system, which resulted in accurate depletion of high-abundance proteins with ease. Our studies provide a convenient and effective method for large-scale depletion of high-abundance proteins and in-depth research in human plasma proteomics.
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, China. xmzhang@fudan.edu.cn
authors.forename: Shaochun
authors.initials: S
authors.lastname: Zhu
authors.forename: Xueyang
authors.initials: X
authors.lastname: Zhang
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gao
authors.forename: Guangfeng
authors.initials: G
authors.lastname: Hong
authors.forename: Guoquan
authors.initials: G
authors.lastname: Yan
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000838:Anions / Q000737:chemistry; D001798:Blood Proteins / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange / Q000191:economics / Q000379:methods*; D006801:Humans; D040901:Proteomics / Q000191:economics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000379:methods; D013997:Time Factors
chemical_list: D000838:Anions; D001798:Blood Proteins