HFIP extraction followed by 2D CTAB/SDS-PAGE separation: a new methodology for protein identification from tissue sections after MALDI mass spectrometry profiling for personalized medicine research.
Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) and profiling technology have become the easiest methods for quickly accessing the protein composition of a tissue area. Unfortunately, the demand for the identification of these proteins remains unmet. To overcome this bottleneck, we combined several strategies to identify the proteins detected via MALDI profiling including on-tissue protein extraction using hexafluoroIsopropanol (1,1,1,3,3,3-hexafluoro-2-propanol, HFIP) coupled with two-dimensional cetyl trimethylammonium bromide/sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D CTAB/SDS-PAGE) for separation followed by trypsin digestion and MALDI-MS analyses for identification. This strategy was compared with an on-tissue bottom-up strategy that we previously developed. The data reflect the complementarity of the approaches. An increase in the number of specific proteins identified has been established. This approach demonstrates the potential of adapted extraction procedures and the combination of parallel identification approaches for personalized medicine applications. The anatomical context provides important insight into identifying biomarkers and may be considered a first step for tissue-based biomarker research, as well as the extemporaneous examination of biopsies during surgery.
1 Laboratoire de Protéomique, Réponse Inflammatoire, Spectrométrie de Masse (PRISM), MALDI Imaging Team, Université de Lille 1 , Cité Scientifique, Lille, France .
Rémi
R
Longuespée
Christophe
C
Tastet
Annie
A
Desmons
Olivier
O
Kerdraon
Robert
R
Day
Isabelle
I
Fournier
Michel
M
Salzet
D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000737:chemistry*; D057285:Precision Medicine; D020543:Proteome / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D057825:Two-Dimensional Difference Gel Electrophoresis
D020543:Proteome