title: An optimised method for the proteomic profiling of full thickness human skin.
abstract: BACKGROUND
  The skin is the largest organ of the human body and is the first line barrier defence against trauma, microbial infiltration and radiation. Skin diseases can be a result of multi-systemic disease or an isolated condition. Due to its proteolysis resistant properties there are relatively few human skin proteomic datasets published compared with other human organs or body fluids. Skin is a challenging tissue to analyse using traditional proteomic techniques due to its high lipid content, insolubility and extensive cross-linking of proteins. This can complicate the isolation and digestion of proteins for analysis using mass spectrometry techniques.
  
  
  RESULTS
  We have optimised a sample preparation procedure to improve solubilisation and mass spectral compatibility of full thickness skin samples. Using this technique, we were able to obtain data for the proteome profile of full thickness human skin using on-line two-dimensional liquid chromatography, followed by ultra-high definition label-free mass spectrometry analysis (UDMS(E)). We were able to identify in excess of 2000 proteins from a full thickness skin sample.
  
  
  CONCLUSIONS
  The adoption of on-line fractionation and optimised acquisition protocols utilising ion mobility separation (IMS) technology has significantly increased the scope for protein identifications ten-fold.
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health, 30 Guilford Street, London, WC1N 1EH UK.
authors.forename: Emily
authors.identifier: 0000-0002-2754-4556
authors.initials: E
authors.lastname: Bliss
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health, 30 Guilford Street, London, WC1N 1EH UK.
authors.forename: Wendy E
authors.initials: WE
authors.lastname: Heywood
authors.affiliation: Histopathology Department, Great Ormond Street Hospital, London, WC1N 3JH UK.
authors.forename: Malika
authors.initials: M
authors.lastname: Benatti
authors.affiliation: Histopathology Department, Great Ormond Street Hospital, London, WC1N 3JH UK.
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Sebire
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health, 30 Guilford Street, London, WC1N 1EH UK.
authors.forename: Kevin
authors.initials: K
authors.lastname: Mills
keywords: Mass spectrometry; Proteomics; Skin profiling; Solubilisation