87818 - Applied Biochemistry

Learning outcomes

Applied Biochemistry has as objectives the study of the theoretical foundations, the instrumental implementations and the main techniques used in cosmetics, in the preparation and analysis of samples for biochemical investigations. The student: o knows the physical and chemical principles underlying the main techniques and instruments employed in biochemical investigations on cosmetic products with reference to the new in vitro tests; -knows the different parameters that may be modified to change biochemical analysis on cosmetic products; o includes a working protocol and the main variables on which to act to optimize it; -can describe the structural elements of the main tools of a biochemical laboratory/cosmetology; -knows the correct terminology used in biochemical methods in cosmetics; -can use the knowledge acquired to evaluate the results of an experiment, identify the problems and optimize it; -knows how to evaluate the possible impact of changes of the parameters in an experiment for the evaluation of corrosion and skin irritation, skin absorption measures and in the main imaging techniques to measure the phototoxicity; o can orientate between the main qualitative and quantitative methodologies, to select the one appropriate to the study of the problem with reference to the new in vitro tests available for the analysis of cosmetic products.

Course contents

Basic principles: Biochemical and molecular biology studies, Quantitative biochemical measurements, Safety in the laboratory.

Cell culture techniques: The cell culture laboratory and equipment, Safety considerations in cell culture, Aseptic techniques and good cell culture practice, Types of animal cell, characteristics and maintenance in culture, Stem cell and bacterial cell culture, Potential use of cell cultures.

Centrifugation: Basic principles of sedimentation, Types, care and safety aspects of centrifuges, Preparative centrifugation, Analytical centrifugation.

Microscopy: The light microscope, Optical sectioning, Imaging living cells and tissues, Measuring cellular dynamics, The electron microscope (EM), Image archiving.

Immunochemical techniques: Making antibodies, Immunoassay formats, Immuno-microscopy, Lateral flow devices, Epitope mapping, Immunoblotting, Fluorescent activated cell sorting , Cell and tissue staining techniques, Immuno-capture polymerase chain reaction (PCR), Immuno-affinity chromatography, Antibody-based biosensors, Therapeutic antibodies.

Protein: structure, purification, characterisation and function analysis.

Electrophoretic techniques: general principles and Support media, Electrophoresis of proteins and of nucleic acids, Capillary electrophoresis, Microchip electrophoresis.

Chromatographic techniques: Principles of chromatography and Chromatographic performance parameters, High-performance liquid chromatography, Adsorption - Partition - Ion-exchange -Molecular (size) exclusion - Affinity and Gas chromatography.

Enzymes: Characteristics and nomenclature, Steady-state kinetics, Analytical methods for the study of enzyme reactions, Active sites and catalytic mechanisms, Control of enzyme activity.

Molecular biology, bioinformatics and basic techniques: Structure of nucleic acids, Genes and genome complexity, Location and packaging of nucleic acids, Functions and Manipulation of nucleic acids, Isolation and separation of nucleic acids, Molecular biology and bioinformatics, Molecular analysis of nucleic acid sequences, Polymerase chain reaction (PCR), Nucleotide sequencing of DNA.

Nods of Recombinant DNA and genetic analysis: Constructing gene libraries, Cloning vectors, Hybridisation and gene probes, Screening gene libraries, Applications of gene cloning, Expression of foreign genes, Analysing genes and gene expression, Analyzing whole genomes, Pharmacogenomics, Molecular biotechnology and applications.

Readings/Bibliography

"Principles and Techniques of Biochemistry and Molecular Biology"

 

Seventh Edition

 

Edited by Keith Wilson and John Wolker

 

Teaching methods

Lectures supported by powerpoint presentations of each topic of the course will be shown

Teaching tools

PC, overhead projector, biological laboratory

Office hours

See the website of Valeria Righi

See the website of Cristiana Caliceti

See the website of Elisa Boschetti