Document Detail


Targeting human serum fucome by an integrated liquid-phase multicolumn platform operating in "cascade" to facilitate comparative mass spectrometric analysis of disease-free and breast cancer sera.
MedLine Citation:
PMID:  23533108     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A fully integrated platform was developed for capturing/fractionating human fucome from disease-free and breast cancer sera. It comprised a multicolumn operated by HPLC pumps and switching valves for the simultaneous depletion of high abundance proteins via affinity-based subtraction and the capturing of fucosylated glycoproteins via lectin affinity chromatography followed by the fractionation of the captured glycoproteins by reversed phase chromatography (RPC). Two lectin columns specific to fucose, namely Aleuria aurantia lectin (AAL) and Lotus tetragonolobus agglutinin (LTA) were utilized. The platform allowed the "cascading" of the serum sample from column-to-column in the liquid phase with no sample manipulation between the various steps. This guaranteed no sample loss and no propagation of experimental biases between the various columns. Finally, the fucome was fractionated by RPC yielding desalted fractions in volatile acetonitrile-rich mobile phase, which after vacuum evaporation were subjected to trypsinolysis for LC-MS/MS analysis. This permitted the identification of the differentially expressed proteins (DEP) in breast cancer serum yielding a broad panel of 35 DEP from the combined LTA and AAL captured proteins and a narrower panel of eight DEP that were commonly differentially expressed in both LTA and AAL fractions, which are considered as more representative of cancer altered fucome.
Authors:
Subhashini Selvaraju; Ziad El Rassi
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2013-04-24
Journal Detail:
Title:  Proteomics     Volume:  13     ISSN:  1615-9861     ISO Abbreviation:  Proteomics     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-06-04     Completed Date:  2013-12-26     Revised Date:  2014-05-07    
Medline Journal Info:
Nlm Unique ID:  101092707     Medline TA:  Proteomics     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1701-13     Citation Subset:  IM    
Copyright Information:
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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MeSH Terms
Descriptor/Qualifier:
Breast Neoplasms / blood*
Case-Control Studies
Chromatography, Affinity
Chromatography, High Pressure Liquid
Chromatography, Reverse-Phase
Female
Fucose / blood*
Glycoproteins / blood*,  isolation & purification
Humans
Lectins / chemistry
Proteome / isolation & purification,  metabolism
Tandem Mass Spectrometry
Tumor Markers, Biological / blood*,  isolation & purification
Grant Support
ID/Acronym/Agency:
1R15GM096286-01/GM/NIGMS NIH HHS; R15 GM096286/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Glycoproteins; 0/Lectins; 0/Proteome; 0/Tumor Markers, Biological; 3713-31-3/Fucose
Comments/Corrections

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