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Proteomics and immunoproteomics

Proteomics and immunoproteomics platform 
The Proteomics and immunoproteomics platform, operated by Porto Conte Ricerche, is currently one of the most comprehensive and state-of-the-art laboratories in Italy.

The services offered by the platform address both systematic proteomics, i.e. identification of the proteins most expressed in a tissue, cell, or cellular element and the building of databases, and differential proteomics, i.e. quantification of differently expressed proteins and identification of molecules involved in physiological and pathophysiological processes.

Its main applications are in the field of innovative diagnostic products:
- search, identification and characterisation of protein markers
- validation of markers identified
- characterisation and purification of antigens for immunisation
- epitope mapping
- peptide design, synthesis and purification
- protein production
- screening of monoclonal antibodies through mass spectrometry
- identification of post-translational modifications
- qualitative and quantitative characterisation of enzyme complexes
- redox switch characterisation, etc.

Contacts
Tonina Roggio
Indirizzo emailroggio@portocontericerche.it
Research and Development Department
Porto Conte Ricerche
S.P. 55 Porto Conte-Capo Caccia, Km 8.400
Loc. Tramariglio
C.P. Santa Maria La Palma
07041 Alghero (SS) - Italy
Phone +39 079 998.400
Fax +39 079 998.567

Equipment
Systems for IEF and 2D PAGE
They enable separation and purification of complex protein mixtures extracted from cells, tissue or other biological specimens.
The 2D technique exploits two independent properties to separate the proteins in two different steps. The first dimension, isoelectric focusing (IEF), exploits the different isoelectric points (pI) of the proteins to be separated. The second dimension, PolyAcrylamide Gel Electrophoresis (SDS-PAGE), discriminates proteins by molecular weight.
The proteins are then highlighted by means of staining on the 2D gel, determining pI and their apparent molecular weight. Excision of the spots identified on the electrophoretic maps and subsequent in situ enzymatic digestion leads through Edman degradation analysis, LC-MS, and MALDI-MS to single protein identification.

Photo Documentation Sytem
Photo documentation system with dark room and high-resolution Peltier cooled CCD camera. The system can also acquire and perform software analysis on images captured by means of fluorescence and chemiluminescence techniques.

NanoHPLC-ESI-Q-Tof
The tandem mass spectrometer, consisting of a Quadrupole-orthogonal acceleration Time Of Flight (Q/oaTOF), is one of the most powerful and versatile tools for structural analysis.
Its innovative geometry offers increased resolution and sensitivity, ensuring greater mass measurement accuracy.
The variety of usable sources (ESI, APCI, Nano Spray) and interfacing with capillary HPLC enable a broad range of applications in the field of proteomics, peptidomics, metabolomics and structural analysis.

MALDI TOF
The instrument, consisting of a TOF (Time Of Flight) analyzer and a MALDI (Matrix Assisted Laser Desorption Ionization) source provides accurate information on the structure and molecular weight of biomolecules such as peptides, proteins, oligonucleotides and carbohydrates, as well as synthetic polymers. Furthermore, through the PMF (Peptide MassFingerprinting) technique, it enables ultra-fast (4 min.) identification of the proteins separated by 2D electrophoresis.
The elute from in situ protein digestion, mixed with the matrix (organic acid), is deposited (1uL) on a steel target and left to crystallise before being transferred to the vacuum system. A laser pulse extracts from the target the molecular ions, which are subsequently accelerated by an electrical field and cross the flight tube, enabling, after appropriate calibration, measurement of molecular weight.

Automated peptide synthesizer
Completely automated system for small and medium-scale peptide synthesis, complete with simultaneous multiple peptide synthesis modules.

EDMAN Sequencer
A completely automated and computerised system with 2 reaction chambers for N-terminal amino acid sequencing by Edman degradation, on samples deposited on glass filter or blotted or covalently bound to a solid support.

Optical Microplate Reader
Microplate reader by UV-Vis spectrophotometry, fluorescence and chemiluminescence.

Semi- and Micro-preparative chromatography systems
Completely automated chromatography systems for semi-micro and micro scale separation and purification of proteins, peptides, molecular probes and oligonucleotides. AKTA Explorer 10 includes autosampler, automated fraction collector, automated column selection valve, and a system for monitoring pH and eluent conductivity. The SMART system for microanalytical and micropreparative chromatography, with multiple wavelength UV detector, and equipped with a microfraction collector, is the ideal tool for highly efficient separation of microquantities of proteins and peptides, sample preparation for LC-ESI-MS, or MALDI analysis, and preconcentration of biomolecules present in small quantities in biological matrixes.