METHODS IN BIOCHEMISTRY
Stampa
Anno immatricolazione
2021/2022
Anno offerta
2021/2022
Normativa
DM270
SSD
BIO/10 (BIOCHIMICA)
Dipartimento
DIPARTIMENTO DI BIOLOGIA E BIOTECNOLOGIE "LAZZARO SPALLANZANI"
Corso di studio
MOLECULAR BIOLOGY AND GENETICS
Curriculum
PERCORSO COMUNE
Anno di corso
Periodo didattico
Primo Semestre (01/10/2021 - 14/01/2022)
Crediti
9
Ore
72 ore di attività frontale
Lingua insegnamento
English
Tipo esame
ORALE
Docente
GUIDETTI GIANNI FRANCESCO (titolare) - 3 CFU
IADAROLA PAOLO - 6 CFU
Prerequisiti
Basic knowledge of biochemistry and molecular biology. Knowledge of
the general concepts about protein structure and function.
Obiettivi formativi
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.
Programma e contenuti
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.
Metodi didattici
Frontal lessons, seminars on specific topics, guided lab tours and description of equipments and facilities for biochemical investigations.
Testi di riferimento
Principles and Techniques of Biochemistry and Molecular Biology. Edited by Wilson and Walker
Modalità verifica apprendimento
Oral presentation of a scientific article. Description of the methods adopted in the paper and discussion of the results.
Altre informazioni
N/A
Obiettivi Agenda 2030 per lo sviluppo sostenibile