66571 - Bioanalytical Proteomics and Interactomics

Learning outcomes

At the end of the course, the student gains the basic knowledge in proteomics (soft impact mass spectrometry technologies and smart sample handling for proteins /peptides quantitative analysis and identification in complex bilogical matrices), interactomics (experimental and bioinformatics tools), methods based on molecular recognition combined with selective and highly detectable probes (Immunoassay, Hybridization reactions, biosensors, FRET,BRET, whole cell biosensors) and Luminescence Molecular imaging: localization and quantification of analytes in single cells and tissue (combined with microscopy), whole organ and living organisms using nanoparticles and recombinant bioluminescent cells.

Course contents

Introduction to proteomics: structural and functional proteomics, strategies for proteomic analysis.

Separative techniques in proteomics: gel electrophoresis (SDS PAGE, 2D PAGE), non-denaturing

gel electrophoresis, capillary electrophoresis, HPLC (reverse phase, ion exchange, size exclusion), multidimensional HPLC.

Mass spectrometry techniques in proteomics: soft ionization techniques, MALDI and ESI ionization sources, quadrupole mass analyzers, time-of-flight mass analyzers (TOF), hybrid analyzers (Q-TOF), mass spectrometers (MALDI/TOF and ESI/Q-TOF), SELDI/TOF analysis.

Protein identification: analysis of intact proteins by ESI/MS and MALDI/MS, in-gel digestion, in-gel digest analysis by MALDI/MS and ESI/MS, database search for protein identification.

Applications of proteomics: proteomics in clinical analysis, “shotgun” proteomics, peptide fingerprint mapping, study of protein complexes, studio di complessi proteici, detection and identification of post-transcriptional protein modifications.

Experimental (Two hybrid systems, FRET, BRET, phage display, microarrays, biosensors) and bioinformatic approaches to predict protein funtion and protein-protein interactions.

Luminescence Molecular Imaging: localization and quantification of analytes in single cells and tissue (combined with microscopy), whole organ and living organisms using nanoparticles and recombinant bioluminescent cells.

In silico prediction of protein-ligand interactions.

Readings/Bibliography

Proteomics in Practice: A Guide to Successful Experimental Design by Westermeier, Naven,Hans-Rudolf Höpker, Wiley-VCH, Weinheim, 2nd ed 2008

Discovering Genomics, Proteomics and Bioinformatics (2nd Edition) by A. Malcolm Campbell and Laurie J. Heyer (Mar 12, 2006)

Teaching methods

The course consists both in lectures dealing about the topics listed in the program and a series of in silico labs (experiments and/or demonstrations concerning common proteomics procedures and application)

Assessment methods

Oral examination

Teaching tools

Video and overhead projector. Slides of all the lectures are available as PowerPoint files.

Office hours

See the website of Elisa Michelini