Learning objectives
KNOWLEDGE AND UNDERSTANDING
At the end of the course, the student is expected to have acquired a systematic knowledge of the main research techniques in biochemistry, including the principles upon which these techniques are based. Furthermore, the student must be familiar with the topics of proteome research, and with the main strategies and techniques employed in proteomics.
CAPACITY TO APPLY KNOWLEDGE AND UNDERSTANDING, AUTONOMY AND COMMUNICATION SKILLS
Besides knowing, understanding and being able to describe the methodologies for the identification, isolation, and structural and functional study of proteins, at the end of the course the student must possess the intellectual tools necessary for the analysis of the experimental results and for their description. For example, in the face of an experimental technique (among those illustrated in the course) that produces a certain result, the student should be able (a) to describe the technique and the data that emerges in appropriate language and (b) analyze critically the experimental data and hypothesize one or more possible interpretations.
The level of understanding of the subject, the mastery of the specific language and the ability to use the acquired knowledge by applying them autonomously in some experimental setting are verified by means of an oral exam.
LEARNING ABILITY
The continuous evolution of scientific research and, in particular, of the biochemical and proteomic methodologies requires a constant update of the pertinet skills. For this reason, the course aims at providing the theoretical basis and the autonomy needed by the students for the continuous upgrade of their know-how as research in the field progresses.
Prerequisites
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Course unit content
Biochemical buffers. 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 (SILAC, ICAT and iTRAQ approaches) and functional proteomics: detailed analysis of some significant examples of proteomic approaches to the study of biological problems. Basic notions of enzymomics and metabolomics.
Full programme
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Bibliography
Notions about the biochemical methods described in the course can be obtained from recent textbooxs such as:
Bonaccorsi di Patti, Contestabile, Di Salvo (2019), Metodologie Biochimiche, Zanichelli
The lectures on proteomics will be based on scientific papers (in English) directly provided by the teacher.
Teaching methods
In the face of the COVID-19 pandemic, the course for the current academic year will consist of online-only lectures. The classes will be streamed synchronously (i.e. the teacher will present his lectures on the Teams platform at the times and dates specified by the teaching calendar). The videos of these classes will remain available to students on the Elly platform (or on another platform indicated by the university) for at least 15 days. In these lectures, the students will be exposed to the main biochemical methods used in protein biochemistry and proteomics.
Assessment methods and criteria
For the current academic year, it is expected that the knowledge acquired, as well as the ability to use it in practice, will be assessed through an oral exam. As long as the COVID-19 emergency continues, the test will be carried out online. During this oral exam, students will be asked to describe in detail some specific techniques covered in the course (this will serve to evaluate the acquired knowledge and understanding of the subject) and above all to recognize how and when to use these techniques. This will serve to assess the ability to apply knowledge and understanding, as well as the autonomy of judgment and the ability to communicate ideas and concepts with clarity and property of language.
The final grade (scale 0-30) will ensue from a balanced evaluation of the various aspects mentioned. The ‘cum laude’ honor will be awarded in the case of students who show excellent knowledge and understanding of the subject, excellent ability to apply them, good independent judgment and fluency in the lexicon of the discipline.
Other information
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