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Enhanced Detection of Low-Abundance Human Plasma Proteins by Integrating Polyethylene Glycol Fractionation and Immunoaffinity Depletion
by
Liu, Zhao
, Zhou, Jian
, Xie, Peng
, Liu, Haipeng
, Yang, Yongtao
, Zhou, Mi
, Fan, Songhua
, Qiao, Rui
, Yu, Jia
in
Abundance
/ Adolescent
/ Adult
/ Bioindicators
/ Biology and Life Sciences
/ Biomarkers
/ Blood plasma
/ Blood proteins
/ Blood Proteins - analysis
/ Blood Proteins - isolation & purification
/ Chemical Fractionation - methods
/ Chromatography
/ Chromatography, Affinity - methods
/ Chromatography, Liquid - methods
/ Collaboration
/ Complexity
/ Depletion
/ Disease
/ Female
/ Fractionation
/ Gene expression
/ Hospitals
/ Humans
/ Laboratories
/ Liquid chromatography
/ Male
/ Mass spectrometry
/ Mass spectroscopy
/ Medical research
/ Medicine and Health Sciences
/ Middle Aged
/ Neurobiology
/ Neurology
/ Neurosciences
/ Peptides
/ Plasma
/ Plasma proteins
/ Polyethylene
/ Polyethylene glycol
/ Polyethylene Glycols - chemistry
/ Proteins
/ Proteomics
/ Research and Analysis Methods
/ Science
/ Spectroscopy
/ Strategy
/ Studies
/ Tandem Mass Spectrometry - methods
/ Young Adult
2016
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Enhanced Detection of Low-Abundance Human Plasma Proteins by Integrating Polyethylene Glycol Fractionation and Immunoaffinity Depletion
by
Liu, Zhao
, Zhou, Jian
, Xie, Peng
, Liu, Haipeng
, Yang, Yongtao
, Zhou, Mi
, Fan, Songhua
, Qiao, Rui
, Yu, Jia
in
Abundance
/ Adolescent
/ Adult
/ Bioindicators
/ Biology and Life Sciences
/ Biomarkers
/ Blood plasma
/ Blood proteins
/ Blood Proteins - analysis
/ Blood Proteins - isolation & purification
/ Chemical Fractionation - methods
/ Chromatography
/ Chromatography, Affinity - methods
/ Chromatography, Liquid - methods
/ Collaboration
/ Complexity
/ Depletion
/ Disease
/ Female
/ Fractionation
/ Gene expression
/ Hospitals
/ Humans
/ Laboratories
/ Liquid chromatography
/ Male
/ Mass spectrometry
/ Mass spectroscopy
/ Medical research
/ Medicine and Health Sciences
/ Middle Aged
/ Neurobiology
/ Neurology
/ Neurosciences
/ Peptides
/ Plasma
/ Plasma proteins
/ Polyethylene
/ Polyethylene glycol
/ Polyethylene Glycols - chemistry
/ Proteins
/ Proteomics
/ Research and Analysis Methods
/ Science
/ Spectroscopy
/ Strategy
/ Studies
/ Tandem Mass Spectrometry - methods
/ Young Adult
2016
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Enhanced Detection of Low-Abundance Human Plasma Proteins by Integrating Polyethylene Glycol Fractionation and Immunoaffinity Depletion
by
Liu, Zhao
, Zhou, Jian
, Xie, Peng
, Liu, Haipeng
, Yang, Yongtao
, Zhou, Mi
, Fan, Songhua
, Qiao, Rui
, Yu, Jia
in
Abundance
/ Adolescent
/ Adult
/ Bioindicators
/ Biology and Life Sciences
/ Biomarkers
/ Blood plasma
/ Blood proteins
/ Blood Proteins - analysis
/ Blood Proteins - isolation & purification
/ Chemical Fractionation - methods
/ Chromatography
/ Chromatography, Affinity - methods
/ Chromatography, Liquid - methods
/ Collaboration
/ Complexity
/ Depletion
/ Disease
/ Female
/ Fractionation
/ Gene expression
/ Hospitals
/ Humans
/ Laboratories
/ Liquid chromatography
/ Male
/ Mass spectrometry
/ Mass spectroscopy
/ Medical research
/ Medicine and Health Sciences
/ Middle Aged
/ Neurobiology
/ Neurology
/ Neurosciences
/ Peptides
/ Plasma
/ Plasma proteins
/ Polyethylene
/ Polyethylene glycol
/ Polyethylene Glycols - chemistry
/ Proteins
/ Proteomics
/ Research and Analysis Methods
/ Science
/ Spectroscopy
/ Strategy
/ Studies
/ Tandem Mass Spectrometry - methods
/ Young Adult
2016
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Enhanced Detection of Low-Abundance Human Plasma Proteins by Integrating Polyethylene Glycol Fractionation and Immunoaffinity Depletion
Journal Article
Enhanced Detection of Low-Abundance Human Plasma Proteins by Integrating Polyethylene Glycol Fractionation and Immunoaffinity Depletion
2016
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Overview
The enormous depth complexity of the human plasma proteome poses a significant challenge for current mass spectrometry-based proteomic technologies in terms of detecting low-level proteins in plasma, which is essential for successful biomarker discovery efforts. Typically, a single-step analytical approach cannot reduce this intrinsic complexity. Current simplex immunodepletion techniques offer limited capacity for detecting low-abundance proteins, and integrated strategies are thus desirable. In this respect, we developed an improved strategy for analyzing the human plasma proteome by integrating polyethylene glycol (PEG) fractionation with immunoaffinity depletion. PEG fractionation of plasma proteins is simple, rapid, efficient, and compatible with a downstream immunodepletion step. Compared with immunodepletion alone, our integrated strategy substantially improved the proteome coverage afforded by PEG fractionation. Coupling this new protocol with liquid chromatography-tandem mass spectrometry, 135 proteins with reported normal concentrations below 100 ng/mL were confidently identified as common low-abundance proteins. A side-by-side comparison indicated that our integrated strategy was increased by average 43.0% in the identification rate of low-abundance proteins, relying on an average 65.8% increase of the corresponding unique peptides. Further investigation demonstrated that this combined strategy could effectively alleviate the signal-suppressive effects of the major high-abundance proteins by affinity depletion, especially with moderate-abundance proteins after incorporating PEG fractionation, thereby greatly enhancing the detection of low-abundance proteins. In sum, the newly developed strategy of incorporating PEG fractionation to immunodepletion methods can potentially aid in the discovery of plasma biomarkers of therapeutic and clinical interest.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ Adult
/ Blood Proteins - isolation & purification
/ Chemical Fractionation - methods
/ Chromatography, Affinity - methods
/ Chromatography, Liquid - methods
/ Disease
/ Female
/ Humans
/ Male
/ Medicine and Health Sciences
/ Peptides
/ Plasma
/ Polyethylene Glycols - chemistry
/ Proteins
/ Research and Analysis Methods
/ Science
/ Strategy
/ Studies
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