title: An automated plasma protein fractionation design: high-throughput perspectives for proteomic analysis.
abstract: BACKGROUND
  Human plasma, representing the most complete record of the individual phenotype, is an appealing sample for proteomics analysis in clinical applications. Up to today, the major obstacle in a proteomics study of plasma is the large dynamic range of protein concentration and the efforts of many researchers focused on the resolution of this important drawback.
  
  
  RESULTS
  In this study, proteins from pooled plasma samples were fractionated according to their chemical characteristics on a home-designed SPE automated platform. The resulting fractions were digested and further resolved by reversed-phase liquid chromatography coupled with MALDI TOF/TOF mass spectrometry. A total of 712 proteins were successfully identified until a concentration level of ng/mL. Pearson correlation coefficient was used to test reproducibility.
  
  
  CONCLUSIONS
  Our multidimensional fractionation approach reduced the analysis time (2 days are enough to process 16 plasma samples filling a 96-well plate) over the conventional gel-electrophoresis or multi-LC column based methods. The robotic processing, avoiding contaminants or lack of sample handling skill, promises highly reproducible specimen analyses (more than 85% Pearson correlation). The automated platform here presented is flexible and easily modulated changing fractioning elements or detectors.
authors.affiliation: Institute of Clinical Physiology-CNR, Via Moruzzi 1, 56124 Pisa, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Boccardi
authors.forename: Silvia
authors.initials: S
authors.lastname: Rocchiccioli
authors.forename: Antonella
authors.initials: A
authors.lastname: Cecchettini
authors.forename: Alberto
authors.initials: A
authors.lastname: Mercatanti
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Citti
mesh_terms: D001798:Blood Proteins / Q000032:analysis*; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D006801:Humans; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001798:Blood Proteins; D020543:Proteome