department of proteomic and microbiology - umons - lab'insight proteomics
DESCRIPTION
description of the laboratory skills (09/03/2010)TRANSCRIPT
18/03/10
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Department of Proteomic and Microbiology (ProtMic)
University of Mons
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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?
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
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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
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
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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
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 [email protected]
10 complementary MS instruments
High experience level in small and large molecules
18/03/10
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Thank you for your aLen>on