title: 2D-electrophoresis and multiplex immunoassay proteomic analysis of different body fluids and cellular components reveal known and novel markers for extended fasting.
abstract: BACKGROUND
  Proteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation.
  
  
  METHODS
  Both a non-targeted (classical 2D-electrophoresis combined with mass spectrometry) as well as a targeted proteomic approach (multiplex immunoassay) were applied to investigate how fasting for 36 h, as compared to 12 h, affects the proteome of platelets, peripheral blood mononuclear cells (PBMC), plasma, urine and saliva collected from ten healthy volunteers.
  
  
  RESULTS
  Between-subject variability was highest in the plasma proteome and lowest in the PBMC proteome. Random Forests analysis performed on the entire dataset revealed that changes in the level of the RhoGDI2 protein in PBMC and plasma ApoA4 levels were the two most obvious biomarkers of an extended fasting. Random Forests (RF) analysis of the multiplex immunoassay data revealed leptin and MMP-3 as biomarkers for extended fasting. However, high between-subject variability may have masked the extended fasting effects in the proteome of the biofluids and blood cells.
  
  
  CONCLUSIONS
  Identification of significantly changed proteins in biofluids and blood cells using a non-targeted approach, together with the outcome of targeted analysis revealed both known and novel markers for a 36 h fasting period, including the cellular proteins RhoGDI2 and CLIC1, and plasma proteins ApoA4, leptin and MMP-3. The PBMC proteome exhibited the lowest between-subject variability and therefore these cells appear to represent the best biosamples for biomarker discovery in human nutrigenomics.
authors: Department of Human Biology, NUTRIM, Maastricht University, Maastricht, The Netherlands.
authors: Freek G
authors: FG
authors: Bouwman
authors: Baukje
authors: B
authors: de Roos
authors: Isabel
authors: I
authors: Rubio-Aliaga
authors: L Katie
authors: LK
authors: Crosley
authors: Susan J
authors: SJ
authors: Duthie
authors: Claus
authors: C
authors: Mayer
authors: Graham
authors: G
authors: Horgan
authors: Abigael C
authors: AC
authors: Polley
authors: Carolin
authors: C
authors: Heim
authors: Susan L M
authors: SL
authors: Coort
authors: Chris T
authors: CT
authors: Evelo
authors: Francis
authors: F
authors: Mulholland
authors: Ian T
authors: IT
authors: Johnson
authors: Ruan M
authors: RM
authors: Elliott
authors: Hannelore
authors: H
authors: Daniel
authors: Edwin C M
authors: EC
authors: Mariman
mesh_terms: D001054:Apolipoproteins A / Q000097:blood; D015415:Biomarkers / Q000378:metabolism; D001773:Blood Cells / Q000378:metabolism*; D001826:Body Fluids / Q000378:metabolism*; D016000:Cluster Analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D005215:Fasting; D020730:Guanine Nucleotide Dissociation Inhibitors / Q000378:metabolism; D006801:Humans; D007118:Immunoassay; D020738:Leptin / Q000378:metabolism; D013058:Mass Spectrometry; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D025341:Principal Component Analysis; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D013997:Time Factors; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D062949:rho Guanine Nucleotide Dissociation Inhibitor beta; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors
chemical_list: C569658:ARHGDIB protein, human; D001054:Apolipoproteins A; D015415:Biomarkers; D020730:Guanine Nucleotide Dissociation Inhibitors; D020738:Leptin; D020543:Proteome; D025521:Tumor Suppressor Proteins; C022205:apolipoprotein A-IV; D062949:rho Guanine Nucleotide Dissociation Inhibitor beta; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; D019278:Matrix Metalloproteinase 3