ANALYTICAL AND CLINICAL PROTEOMICS (2012/2013)

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Course code
4S000680
Credits
6
Coordinator
Daniela Cecconi
Other available courses
Other available courses
    Academic sector
    CHIM/01 - ANALYTICAL CHEMISTRY
    Language of instruction
    Italian
    Teaching is organised as follows:
    Activity Credits Period Academic staff Timetable
    Teoria 5 II semestre Daniela Cecconi
    Laboratorio 1 II semestre Daniela Cecconi

    Lesson timetable

    Learning outcomes

    Clinical proteomics is the science that aims to develop technologies for the treatment of patients through the analysis of proteomes of healthy and sick cells. This research area aimed to identify proteins that can be used as early markers in blood, urine, or tissues of cancer patients, or that may predict response to therapy or the relapse after treatment.

    Syllabus

    PROTEOMICS

    Analytical proteomics (LM9+LM7):

    1. Introduction.

    2. Sample preparation.

    3. Gel-based separation.

    4. 2D gel Image analysis.

    5. Gel-free separation.

    6. Pre-fractionation of low abundance proteins.

    7. Analysis of proteins/peptides by Mass Spectrometry.

    8. Analysis of proteins/peptides by Tandem Mass Spectrometry.

    9. Analysis of mass spectra and database search for protein identification.

    10. Differential and Quantitative proteomics.

    11. Characterization of post-translational modifications.

    12. Proteomics in agro-food and biomedical areas.


    Clinical proteomics (LM9):

    13. Application of Proteomics in Cancer Research.

    14. Proteomics and lymphomas (mantle cell lymphoma and splenic lymphoma).

    15. Proteomics and stem cells. Study of mechanism of action of new drugs.

    16. From iron-diseases to hypertension: application of clinical proteomics.

    17. Hemochromatosis and Intrauterine growth retardation

    18. Cardiac diseases.

    19. Autoimmune disease.

    20. Bacterial and viral infections.




    Laboratory (1 CFU)

    Day 1:
    Rehydration of 7 cm IPG with protein extract
    Preparation of buffers and solutions for 2-DE analysis
    Polymerization of 12% T gel
    Isoelectrofocusing by using PROTEAN® IEF Cell (Bio-Rad)


    Day 2:
    IPG strip equilibration
    Interfacing with the second dimension
    Second dimension by Mini-PROTEAN® 3 Dodeca™ Cell (Bio-Rad)
    Staining of 2-DE maps with Coomassie and/or SyproRuby

    Day 3:
    Destaining and acquisition of images by VersaDoc™ MP 1000 system (Bio-Rad)
    Image analysis by PDQuest software (Bio-Rad) (classroom gamma)
    Analysis of mass spectra and database searching with Mascot

    Assessment methods and criteria

    written

    Mandatory attendance to laboratory.

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