APPLIED BIOCHEMISTRY
Written test with multiple choice questions, and open questions
The course aims to raise knowledge of the principles of the different methodologies for the identification, characterization and purification of biological macromolecules, and for their structural and functional characterization in biochemistry. The knowledge of the investigation procedures on the structure, function of nucleic acids, gene expression and pharmacogenomics will also be deepened. Will be explanied the main methods of investigation in proteomics and metabolomics, with particular reference to mass spectrometry, applied to pharmaceutical and clinical sciences.
At the end of the training activity the student should be able to: to understand the principles of the main biochemical techniques, understand an experimental design based on the biochemical techniques addressed to deal with common problems related to DNA and or biomolecules and their characterization; to know how to define which biochemical technique it is more suitable to obtain information about a biomolecule or a substitute or a derivative or a biotechnological product; to link the knowledge of the other teachings, in particular in the field of pharmacogenomics, to analyze the mechanism of action of drugs from a molecular and biochemical perspective, to be able to read and interpret scientific publications on the subject.
Study of the analytical methodologies applied to biochemistry, Quantitative biochemical measurements, Safety in the laboratory, Centrifugation (Basic principles of sedimentation, preparative and analytical centrifugation, types, care and safety aspects of centrifuges).
Molecular biology and basic techniques (Structure of nucleic acids, Genes and genome complexity, Functions of nucleic acids, The manipulation of nucleic acids – basic tools and techniques, Isolation and separation of nucleic acids, nucleic acid sequences, The polymerase chain reaction (PCR), Nucleotide sequencing of DNA, Analysing genes and gene expression, Analysing whole genomes, Pharmacogenomics.
Immunochemical techniques: Immunoassay formats, Immunoblotting, Therapeutic antibodies.
Protein structure, purification, characterisation and function analysis, ionic properties of amino acids and proteins.
Spectroscopic techniques: (Ultraviolet and visible light spectroscopy, Fluorescence spectroscopy, Luminometry, Light scattering , Atomic spectroscopy, Infrared and Raman spectroscopy, Electron paramagnetic resonance, Nuclear magnetic resonance, X-ray diffraction.
Chromatographic techniques : (Principles of chromatography, Chromatographic performance parameters,
High-performance liquid chromatography, Adsorption chromatography, Partition chromatography, Ion-exchange chromatography, Molecular (size) exclusion chromatography, Affinity chromatography, Gas chromatography.
Mass spectrometric techniques (Introduction, Ionisation, Mass analysers, Detectors, Structural information by tandem mass spectrometry, Analysing protein complexes, Computing and database analysis
Electrophoretic techniques : (General principles, Electrophoresis of proteins, Electrophoresis of nucleic acids, Capillary electrophoresis,
Omic sciences: genomics, transcriptomics, proteomics, glycomics, metabolomics, lipidomics. Definition and analysis of the Proteome, extraction of the protein from the sample, - separation by one-dimensional and or by two-dimensional electrophoresis, isoelectrofocusing, image analysis of the patterns (peptide mass fingerprint), identification of proteins in mass spectrometry.
Definition and analysis of the metabolome, metabolites (sugars, nucleotides, amino acids, lipids), approaches used in metabolomics (target analysis and non-target analysis)
Metabolomics applications in the biomedical field of: toxicology, oncology, vascular disorders.
Molecular Biology and Biotechnology, 5th Edition Edited by John M Walker and Ralph Rapley
“Metabolomics The Frontier of Systems Biology” - “Tomita – Nishioka” Springer
Clinical Biochemistry, N. Amhed, ed. Oxford University Press(UK).
Lecture notes, slides and scientific papers selected by the Teacher
Frontal lessons: based on simultaneous work with all students in the entire classroom by media system presentation.
The course is intended for the students in the Pharmacy and Chemistry and Pharmaceutical Technologies Course. The course will be held as an intensive course, distributed on one semester.