Searching biomarker candidates in serum using multidimensional native chromatography I. Enhanced separation method

The microplate-based method developed by our group for non-denaturing multidimensional proteome separation was improved on using improved column arrays and a newly developed robot. Currently size exclusion, anion exchange and lectin affinity chromatography are combined orthogonally. Different sample...

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Published inJournal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 875; no. 2; pp. 567 - 572
Main Authors KREUSCH, Stefan, SCHULZE, Margarete, CUMME, Gerhard A, DITZE, Günter, MOORE, Thomas, RHODE, Heidrun
Format Journal Article
LanguageEnglish
Published Amsterdam Elsevier 15.11.2008
Subjects
Analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Biomarkers - blood
Blood Proteins - analysis
Chromatography - methods
Chromatography, Affinity - methods
Chromatography, Gel - methods
Chromatography, High Pressure Liquid - methods
Chromatography, Ion Exchange - methods
Databases, Protein
Fundamental and applied biological sciences. Psychology
General pharmacology
Humans
Medical sciences
Peptides - analysis
Pharmacology. Drug treatments
Proteomics - instrumentation
Proteomics - methods
Reproducibility of Results
Robotics - instrumentation
Spectrometry, Mass, Electrospray Ionization - methods
Tandem Mass Spectrometry - methods
Human
Multidimensional chromatography
Biological fluid
Separation method
Human serum proteome
Microtiter plate
Biological marker
Microplate
Proteome
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Summary:The microplate-based method developed by our group for non-denaturing multidimensional proteome separation was improved on using improved column arrays and a newly developed robot. Currently size exclusion, anion exchange and lectin affinity chromatography are combined orthogonally. Different samples run simultaneously to enhance reliability of intercomparison. LC-ESI (electro-spray ionization) MS/MS analysis of selected fractions identified 32,288 peptides matching 2,669 serum proteins. The present contribution (I) shows the characteristics of the method, whereas "prove of principle" by applying it to search for biomarker candidates with model diseases is reported in an accompanying paper (II).
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2008.09.040