BIOCHEMICAL AND PROTEOMICS METHODS
Learning outcomes of the course unit
In this course, the students will receive a systematic preparation on the main techniques used in protein biochemistry and proteomics. Students will become familiar with the methods for the identification, isolation and characterization of proteins, as well as with the intellectual tools needed to analyze the results and to describe them. Furthermore, they will be informed about the themes, the strategies and major techniques employed in the field of proteomics. The acquired knowledge and understanding will be assessed through written exams, in which students will be asked to describe specific techniques in detail and above all to recognize how and when to employ such techniques.
Course contents summary
Extraction and purification of proteins.
Chromatographic techniques: gel filtration chromatography, ion exchange, hydrophobic exchange and affinity chromatography.
Protein electrophoresis: native, SDS-PAGE, isoelectrofocusing, 2D-PAGE.
Immunochemical techniques: immunoprecipitation, ELISA, Western blotting.
Optical spectroscopy techniques and their applications: absorption, spectrofluorometry and circular dichroism.
Basic notions of nuclear magnetic resonance spectroscopy (NMR).
Basic notions on the use of radioisotopes.
Enzymatic assay methods. Continuous and discontinuous assays. Methods based on coupled reactions. Activity Units.
Mass spectrometry: ionization systems and analytical methods (magnetic deflection detectors, quadrupole, flight time, orbitrap, tandem detectors).
Proteomics, general concepts.
Systemic proteomics and methodological basics of the proteomic approach: mono- and bi-dimensional proteins of electrophoresis; Mass spectrometry coupled with the analysis of databases. Proteomic study of post-translational modifications.
Differential proteomics and functional proteomics: detailed analysis of some significant examples of proteomic approaches to the study of biological problems. Basic notions of enzymomics and metabolomics.
Notions about the biochemical methods described in the course can be obtained from texts such as:
Bonaccorsi, Contestabile, Di Salvo, Metodologie Biochimiche, Casa Editrice Ambrosiana; Reed, Holmes, Weyers & Jones, Metodologie di base per le scienze biomolecolari,
Zanichelli. The lectures on proteomics will be based on scientific papers (in english) directly provided by the teacher.
The course will be based on formal classroom lectures, in which the students will be exposed to the main biochemical methods used in protein biochemistry and proteomics.
Assessment methods and criteria
The knowledge and understanding acquired by the student, as well as her/his ability to apply such a knowledge in practice, will be assessed through a written test. The test will encompass a section with 15 multiple-choice quizzes and a section including two open questions. The multiple-choice quizzes will mainly serve to assess the acquired knowledge and understanding. As for open questions, the student will be asked to describe specific techniques in detail and above all to recognize how and when to employ such techniques . This will serve to assess the ability to apply knowledge and understanding, as well as the capacity of making judgements and of communicating ideas and concepts with clarity and property of language.
In the multiple-choices section, students will be awarded one point for each exact answer (maximum score in the section: 15). As for the open questions, each of them will be attributed a score between 0 and 7.5 (maximum score in the section: 15).
The final grade (scale 0-30) will represent the sum of the scores for the two sections. The 'cum laude' honor is assigned when both sections achieve maximum score and the candidate shows fluency the lexicon of the disicpline.