The course aims to provide a thorough overview of the biochemical methods used in experimental research, medicine and in the general field of biotechnology. The main focus will be on the production and charaterization of proteins (e.g. antibodies, natural and recombinant enzymes, hormons) and their use in diagnosis, therapy and industrial applications. The students will acquire the ability to provide a complete description and practical examples of several common techniques used in the biochemical investigations and to critically discuss the most important aspects of experimental biochemistry.
Course Prerequisites
Basic knowledge of biochemistry. Knowledge of the general concepts about protein structure and function.
Teaching Methods
Frontal lessons, seminars on specific topics, and description of equipment and facilities for biochemical investigations.
Additional support and teaching aids will be guaranteed exclusively to students belonging to specific categories defined by the University governance (working students, disabled, caregivers, etc.), who will not be able to attend the lessons in presence.
Assessment Methods
Presentation and discussion of a scientific article. Description of the methods adopted in the paper and discussion of the results. The score will be expressed on a scale of 30 point. Several exam dates will be scheduled in each semester, in particular during the summer session (in June-July), in order to facilitate the admission to the final Laurea exam for the students enrolled in the second year of the Master Degree.
Texts
Metodologie biochimiche e biomolecolari (Maccarrone) - Zanichelli Metodologie biochimiche (Bonaccorsi di Patti, Contestabile, Di Salvo) - Zanichelli
Contents
DNA recombinant technology: cloning, mutagenesis and expression of recombinant proteins. PCR and real-time PCR. Methods for extraction and purification of proteins from animal/vegetal tissues and cultured cells. Procedures for primary purification of proteins. Theoretical principles and practical protocols of the most common chromatographic approaches. Electrophoretic techniques: monodimensional (1-DE) and bidimensional (2-DE) electrophoresis. Spectroscopic techniques for the characterization of proteins. Mass Spectrometry (MS). Radioisotopes: principles, manipulation, types of emission, detection, and measurement of radiation. Carbon dating. Use of radioisotopes in biochemistry. Immunochemical techniques: production and purification of polyclonal and monoclonal antibodies and their application in biochemistry and medicine. Enzyme-linked immunosorbent assay (ELISA). Biochemistry of antigenic and molecular tests to assay viral infections. Immunofluorescence analyses: fluorescence and confocal microscopy and flow cytometry.
