Abstract：
:Novel biomarker verification assays are urgently required to improve the efficiency of biomarker development. Benefitting from lower development costs, multiple reaction monitoring (MRM) has been used for biomarker verification as an alternative to immunoassay. However, in general MRM analysis, only one sample can be quantified in a single experiment, which restricts its application. Here, a Hyperplex-MRM quantification approach, which combined mTRAQ for absolute quantification and iTRAQ for relative quantification, was developed to increase the throughput of biomarker verification. In this strategy, equal amounts of internal standard peptides were labeled with mTRAQ reagents Δ0 and Δ8, respectively, as double references, while 4-plex iTRAQ reagents were used to label four different samples as an alternative to mTRAQ Δ4. From the MRM trace and MS/MS spectrum, total amounts and relative ratios of target proteins/peptides of four samples could be acquired simultaneously. Accordingly, absolute amounts of target proteins/peptides in four different samples could be achieved in a single run. In addition, double references were used to increase the reliability of the quantification results. Using this approach, three biomarker candidates, ademosylhomocysteinase (AHCY), cathepsin D (CTSD), and lysozyme C (LYZ), were successfully quantified in colorectal cancer (CRC) tissue specimens of different stages with high accuracy, sensitivity, and reproducibility. To summarize, we demonstrated a promising quantification method for high-throughput verification of biomarker candidates.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Yin HR,Zhang L,Xie LQ,Huang LY,Xu Y,Cai SJ,Yang PY,Lu HJdoi
10.1021/pr4005025subject
Has Abstractpub_date
2013-09-06 00:00:00pages
3912-9issue
9eissn
1535-3893issn
1535-3907journal_volume
12pub_type
杂志文章abstract：:Moving from macroscale preparative systems in proteomics to micro- and nanotechnologies offers researchers the ability to deeply profile smaller numbers of cells that are more likely to be encountered in clinical settings. Herein a recently developed microscale proteomic method, microdroplet processing in one pot for ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00629
更新日期：2021-01-25 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00654
更新日期：2020-11-06 00:00:00
abstract：:The Chromosome-Centric Human Proteome Project (C-HPP) aims to identify "missing" proteins in the neXtProt knowledgebase. We present an in-depth proteomics analysis of the human sperm proteome to identify testis-enriched missing proteins. Using protein extraction procedures and LC-MS/MS analysis, we detected 235 protei...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00400
更新日期：2016-11-04 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00488
更新日期：2015-09-04 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00371
更新日期：2020-09-04 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060382t
更新日期：2007-02-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr050279o
更新日期：2006-02-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr801048z
更新日期：2009-05-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00189
更新日期：2020-08-07 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00135
更新日期：2018-09-07 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900399q
更新日期：2009-10-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr060152e
更新日期：2006-09-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr400731p
更新日期：2014-02-07 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300953k
更新日期：2013-02-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500202e
更新日期：2014-08-01 00:00:00
abstract：:Triply and doubly charged iTRAQ ( isobaric tagging for relative and absolute quantitation) labeled peptide cations from a tryptic peptide mixture of bovine carbonic anhydrase II were subjected to electron transfer ion/ion reactions to investigate the effect of charge bearing modifications associated with iTRAQ on the ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8001113
更新日期：2008-09-01 00:00:00
abstract：:Biomarkers for colorectal cancer (CRC) early diagnosis are currently lacking. The purpose of this study was to interpret molecular events in the early stage of CRC that may bring about new biomarkers for early diagnosis. Methylation isotope labeling assistant gel-enhanced liquid chromatography-mass spectrometry (GeLC-...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100406z
更新日期：2010-09-03 00:00:00
abstract：:Human milk oligosaccharides (HMOs) perform a number of functions including serving as prebiotics to stimulate the growth of beneficial intestinal bacteria, as receptor analogues to inhibit binding of pathogens, and as substances that promote postnatal brain development. There is further evidence that HMOs participate ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100362f
更新日期：2010-08-06 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2013-05-03 00:00:00
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journal_title：Journal of proteome research
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journal_title：Journal of proteome research
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00672
更新日期：2020-07-02 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900597s
更新日期：2009-12-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2019-12-06 00:00:00
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journal_title：Journal of proteome research
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更新日期：2010-04-05 00:00:00
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journal_title：Journal of proteome research
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更新日期：2014-01-03 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2010-06-04 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
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更新日期：2004-01-01 00:00:00
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journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00144
更新日期：2018-08-03 00:00:00
