An optimised method for the proteomic profiling of full thickness human skin

Biological Procedures Online

Methodology

Emily Bliss¹  Wendy E. Heywood¹  Kevin Mills¹  Neil J. Sebire²  Malika Benatti² 

[1] Centre for Translational Omics, UCL Institute of Child Health, 30 Guilford Street, WC1N 1EH, London, UK;Histopathology Department, Great Ormond Street Hospital, WC1N 3JH, London, UK;

关键词: Proteomics;    Skin profiling;    Mass spectrometry;    Solubilisation;   

DOI  :  10.1186/s12575-016-0045-y

 received in 2016-03-16, accepted in 2016-06-24,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundThe 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.ResultsWe 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 (UDMSE). We were able to identify in excess of 2000 proteins from a full thickness skin sample.ConclusionsThe 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.

【 授权许可】

CC BY   
© The Author(s). 2016