Proteomics and Metabolomics

The Proteomics and Metabolomics Core Facility at the Center of Advanced Studies and Technologies (CAST) provides high sensitivity proteomics and metabolomics (including metallomics and lipidomics) analyses using state-of-the-art mass spectrometry instruments and methods.

The mission of the facility is to provide a stimulating and appealing environment able to manage and develop mass spectrometry tools and know-how that exceed the normal resources of individual research programs. Moreover, we aim to provide an environment rich in expertise and educational resources, to develop and establish novel methods using our instrumentation, and finally to foster collaboration within UdA scientific community and beyond.

Proteomics Applications

Approaches able to elucidate protein identity and their functions and interactions in cells, biological fluids, conditioned media, sub-cellular recipients or tissues include:

-LC-MS/MS protein identification from digested samples obtained from gel band/spot, chromatographic fractions, immunoprecipitated fractions, or complex samples (biofluids, tissues, enriched exosomes and extracellular vesicles).

-Relative quantification of proteome by label free, using high resolution and high mass accuracy LC-MS and LC-MS/MS.

-LC-MS/MS identification of protein post-translational modifications (PTMs).

-LC-MS Top down high-resolution accurate mass ESI-MS analysis of intact proteins.

-MALDI-MS (MS/MS) protein and peptide analysis/profiling.

-MALDI-TOF-MS IMAGING proteomics on tissue samples.

- Quantitative metallomics approaches in different biological matrices (urine, plasma, cell media, food, drinks...) for several quantifiable elements (Arsenic, Beryllium, Cadmium, Cobalt, Chrome, Iron, Iodine, Manganese, Nickel, Lead, Platinum, Copper, Zinc)

Metabolomics Applications

Our advanced metabolomics methods allow us to perform detection and quantification of various classes of molecules in a variety of biological matrices in targeted and untargeted mode. The available technologies include:

- Targeted quantitative methods for the analysis of polar metabolites, steroids, drugs and lipids.

- LC-MS and LC-MS/MS high resolution mass spectrometer untargeted methods for relative quantitation of polar metabolite and lipidomics profiling.

- MALDI-TLC-MAGING of polar lipids.

- MALDI-TOF- MS-IMAGING of polar lipids

INFO Access

The Proteomics and Metabolomics shared resource provides high sensitivity analyses using state-of-the-art mass spectrometry instruments and methods. Due to the high specificity and complexity of the instrumentations, for each application the collaboration of the facility Staff (with the know-how and expertise needed to manage and develop mass spectrometry tools) is required.

• Thermo Scientific Orbitrap Fusion Tribrid Mass Spectrometer

Equipped with:

• U3000 NANO RSLCNANO SYS FOR EASY-SPRAY
• UltiMate 3000 RSLCnano for Easy-Spray

• MALDI TOF/TOF autoflex speed Bruker Daltonics

Equipped with:

• ImagePrep System Device for IMAGING setup (Bruker Daltonics)

• GC-MS (6890N GC equipped with a 7863 series autosampler and coupled with a 5973N mass spectrometer) Agilent Tecnologies.
• ICP-MS (7500A ICP quadrupolar mass spectrometer) Agilent Technologies
• HPLC-ESI Triple Quadrupole MS (TQUATTRO Ultima Pt, Micromass-WATERS

• Software
  
  • MaxQuant (Max Planck Institute of Biochemistry, Martinsried, Germany)
  • Perseus (statistical tools for high-dimensional omics data analysis)
  • Ingenuity Pathways Analysis “IPA” (integration, and functional interpretation of data derived from ‘omics experiments)
  • MetaboAnalyst (statistical, functional and integrative analysis of metabolomics data)