description of the laboratory skills (09/03/2010)

1. 18/03/10
1
Department of Proteomic and Microbiology
(ProtMic)
University of Mons
2
University of Liège ‐ 9 March 2010
Proteins expression/
purifica>on
Valida>on
Protein
characteriza>on
Proteome
Prefrac>ona>on
Proteomic strategy
op>miza>on
Project concept at ProtMic
Proteins target
discovery
 Experimental design
 Global analysis vs. Targeted analysis
 Gel based approach vs. gel free
 Label‐free vs. Label based quan>fica>on
 Prefrac>ona>on needed?

2. 18/03/10
2
3
University of Liège ‐ 9 March 2010
Proteins expression/
purifica>on
Valida>on
Protein
characteriza>on
Proteome
Prefrac>ona>on
Proteomic strategy
op>miza>on
Project concept at ProtMic
Proteins target
discovery
Targeted analyses Global analyses
 TiO2 phopshopep=des enrichment
 Surface exposed proteins purifica=on
 …
Top down or boLom up approaches
 IMAC‐based proteome prefrac=ona=on
 HPLC and FPLC
 Liquid isoelectrofocusing (protein or pep=des level)
 …
New DPC chips
4
University of Liège ‐ 9 March 2010
Proteins expression/
purifica>on
Valida>on
Protein
characteriza>on
Proteome
Prefrac>ona>on
Proteomic strategy
op>miza>on
Project concept at ProtMic
Proteins target
discovery
Gel‐based proteomic (Immunoproteomic) Gel free differen>al proteomic

3. 18/03/10
3
5
University of Liège ‐ 9 March 2010
Proteins expression/
purifica>on
Valida>on
Protein
characteriza>on
Proteome
Prefrac>ona>on
Proteomic strategy
op>miza>on
Project concept at ProtMic
Proteins target
discovery
 Bacterial expression
 Tag-based purification
 Untagged proteins purification
6
University of Liège ‐ 9 March 2010
Proteins expression/
purifica>on
Valida>on
Protein
characteriza>on
Proteome
Prefrac>ona>on
Proteomic strategy
op>miza>on
Project concept at ProtMic
Proteins target
discovery
 An=body‐based valida=on
 MS‐based valida=on

4. 18/03/10
4
University of Liège ‐ 9 March 2010
Proteins expression/
purifica>on
Valida>on
Protein
characteriza>on
Proteome
Prefrac>ona>on
Proteomic strategy
op>miza>on
Project concept at ProtMic
Proteins target
discovery
 Enzyme ac>vity
 Structural characteriza>on
‐ MS based characteriza>on
‐ Amino acid analysis
‐ Chromatography analysis (HPLC,…)
‐ Spectrophotometry characteriza>on
‐ Purity
‐ Post transla-onal modifica-on
studies (Phosphoryla-on , acetyla-on,
formyla-on, determina-on of S‐S link….)
‐ Protein‐molecule interac-ons
‐ Conforma-onal studies
University of Liège ‐ 9 March 2010
Project concept at ProtMic
Interdisciplinary center of mass spectrometry : CISMA
« A new academic tool dedicated to small molecules and
synthe>c or biological polymers”
open to the academic and industrial partners
Scien=fic boarding : Dr. P. Gerbaux
Prof. R. Muller
Prof. P. Dubois
Prof. M. De Meyer
Prof. R. Sneyders
Director : Prof. R. WaYez
Ruddy.waYez@umons.ac.be
10 complementary MS
instruments
High experience level in
small and large molecules

5. 18/03/10
5
Thank you for your aLen>on