Institut Claudius Regaud

Dr Ben Allal, manager

Presentation

The "protein profile expression" platform of the Claudius Regaud Institute is part of the "Interactions and Expression of Protein Profiles" of the IFR 150, it is itself connected to the protein platform of Génopole implanted within the IPBS. This platform offers a set of resources, abilities and advices in the field of expression of protein profile via the SELDI-TOF technology.

Technology

The SELDI-TOF technology (Surface Enhanced Laser Desorption Ionisation - Time Of Flight) allows the obtentaining of mass protein profiles from very low complex samples quantities rather they are biologic or not (blood, urine, liquid effusion, lysat, cells…). This technology combines the affinity ofchromatography, in order to capture sample proteins in a selective manner on varied chemical surfaces, and the mass spectrometry in time of flight, to assign a molecular mass to each proteins pace retained. Different surfaces / strips are available, allowing the capture and analysis of sub-population proteins based on their physio-chemical properties (hydrophobic proteins, anionic or cationic proteins, binding metal proteins…).

Applications

Profiling studies constitute the major application of this technology. These studies help with the comparison of sample protein profiles (serum, plasma, urine, cell extracts, etc…) in order to highlight the eventual expression differences which can represent possible biomarkers. The use of different chemical surfaces associated with stringency washing can then allow the establishment of a strategy for the purification of interest proteins. The SELDI-TOF technology can also be benieficial as part of studies related to molecular interactions, via the use of pre-activated surfaces, but also for the verification of the purity of the sample during the different purification phases etc…

The platform provides:

• Protein profiles of complex biological samples (Blood, urine, effusion of liquid, cell lysat via the SELDi-TOF (Surface Enhanced Laser Desorption Ionisation - Time Of Flight) technique which combines the chromatographic affinity (to capture protein samples in a selective manner via varied chemistry surfaces) and the mass spectrometry in time of flight (to assign a molecular mass to each protein space retained) ;
• Electrophoresis 1/2D ;
• Gel analysis 1D/2D ;
• Advises, methodological help, and the application orientation of different techniques in proteomics ;
• The advise and help related to the conception and the implementation of experiences ;
• The advise and help related to purification ;
• The advise and help related to identification ;
• The advice and help related to the composition of reports and publication, as well as the valorisation of the results.

Equipments

• Biomek 2000 (Beckman Coulter) for the treatment of samples and preparation of strips.

• PCS 4000 Commercial edition (ProteinChip Reader Biorad): acquisition of protein profiles.

• Densitometer GS800 PC 1D/ 2D (BioRad)

• Software ProteinChip Data Manager : data analysis

• Software QuantityOne and PDQUEST : analysis of gel 1D and 2D.