These authors contributed equally to this work.
Microproteomics by liquid extraction surface analysis: Application to FFPE tissue to study the fimbria region of tubo-ovarian cancer
Article first published online: 6 MAR 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PROTEOMICS - Clinical Applications
Special Issue: Proteomic Analysis of Formalin Fixed Tissue
Volume 7, Issue 3-4, pages 234–240, April 2013
How to Cite
Wisztorski, M., Fatou, B., Franck, J., Desmons, A., Farré, I., Leblanc, E., Fournier, I. and Salzet, M. (2013), Microproteomics by liquid extraction surface analysis: Application to FFPE tissue to study the fimbria region of tubo-ovarian cancer. Prot. Clin. Appl., 7: 234–240. doi: 10.1002/prca.201200070
Colour Online: See the article online to view Fig. 3 in colour.
- Issue published online: 14 APR 2013
- Article first published online: 6 MAR 2013
- Accepted manuscript online: 21 JAN 2013 10:49AM EST
- Manuscript Accepted: 17 DEC 2012
- Manuscript Revised: 26 OCT 2012
- Manuscript Received: 30 JUL 2012
- “Comité du Nord de la Ligue Nationale contre le Cancer”
- University of Lille 1, French Region “Nord Pas de Calais”
- ARCIR FEDER funds for FT MS instrument
- Ministère de l'Enseignement Supérieur et de la Recherche
- Institut Universitaire de France (I. Fournier) and SIRIC ONCO Lille
- Liquid extraction surface analysis;
- Shotgun proteomics;
- Signaling pathway;
- Tubo-ovarian cancer
We have developed a new method for rapid analysis of a specific region on formalin fixed and paraffin embedded (FFPE) tissue sections. This method combines advantages of direct tissue MS analysis keeping histological information and conventional proteomics approaches for confident identification of proteins in complex sample.
After histological annotation, heat-induced antigen retrieval is performed on FFPE tissue. Using a chemical inkjet printer, trypsin is deposited on discrete regions of less than 1 mm2. After protein digestion, a liquid extraction is performed to retrieve all the peptides. Data coming from identification of proteins in cancer and benign region are compared.
In total, 3649 unique peptides were identified (with a peptide strict false discovery rate less than 1%) corresponding to 983 and 792 nonredundant protein groups identified in benign and cancer region, respectively. A total of 123 protein groups are found only in cancer region and 315 are specific to the benign part. From these data, it has been possible to obtain different important signaling pathways involved in cancer processes and some proteins already known as biomarkers.
Conclusions and clinical relevance
This new approach using a combination of localized on-tissue protein digestion and liquid microextraction followed by LC-MS/MS analysis is useful for advancing our understanding of cancer biology. It is a rapid and innovative technique that will contribute positively to clinical proteomics.