Enrollment year
2021/2022
Academic discipline
BIO/10 (BIOCHEMISTRY)
Department
DEPARTMENT OF BIOLOGY AND BIOTECHNOLOGY "LAZZARO SPALLANZANI"
Curriculum
PERCORSO COMUNE
Period
1st semester (01/10/2021 - 14/01/2022)
Lesson hours
72 lesson hours
Prerequisites
Basic knowledge of biochemistry and molecular biology. Knowledge of
the general concepts about protein structure and function.
Learning outcomes
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 contents
Methods for extraction and purification of proteins from animal/vegetal tissues and cultured cells. Procedures typically used in industry/research laboratories for primary purification of proteins. Theoretical principles and practical protocols of the most common chromatographic approaches: adsorption; partition; ion-exchange (IEC); gel-filtration (SEC); affinity; hydrophobic-interaction (HIC); perfusion; gas-chromatography (GC) and supercritical-fluid chromatography (SFC).Chromatofocusing; high performance liquid chromatography (HPLC); fast protein liquid chromatography (FPLC) and principles of green chromatography.
Electrophoretic techniques: analytical and preparative monodimensional (1-DE) electrophoresis of proteins in their native and denaturated state. Sodium-dodecyl-sulfate gel electrophoresis (SDS-PAGE). Isoelectrofocusing (IEF). Two-dimensional (2-DE) electrophoresis: application to proteomic studies. Capillary Zone electrophoresis (CZE) and Micellar Electrokinetic Chromatography (MEKC).
Spectroscopic techniques for the qualitative/quantitative characterization of proteins.
Adsorption spectroscopy: ultraviolet (U.V.); Visible; Infrared (IR); Nuclear Magnetic Resonance (NMR); Electron Spin Resonance (ESR). Emission spectroscopy: fluorescence and phosphorescence. Mass Spectrometry (MS).
Radioisotopes: principles, manipulation, types of emission, detection and measurement of radiation. Use of radioisotopes in biochemistry. Immunochemical techniques: production and purification of polyclonal and monoclonal antibodies and their application in biochemistry. Radioimmuno assay (RIA) and enzyme-linked immunosorbent assay (ELISA): applications in industry, clinical biochemistry and environmental monitoring.
Teaching methods
Frontal lessons, seminars on specific topics, guided lab tours and description of equipments and facilities for biochemical investigations.
Reccomended or required readings
Principles and Techniques of Biochemistry and Molecular Biology. Edited by Wilson and Walker
Assessment methods
Oral presentation of a scientific article. Description of the methods adopted in the paper and discussion of the results.
Sustainable development goals - Agenda 2030
Università di Pavia - Offerta formativa