00062 - Applied Biochemistry

Learning outcomes

The aim of this course is to give a basic knowledge about the major techniques and principles which underlie biochemical experiments and clinical analyses.

Course contents

General principles of biochemical investigations: methods for investigating metabolism, in vivo models, in vitro models, primary cell cultures and cell lines.

Measurements of enzyme activity: enzyme steady state kinetics, enzyme inhibitors, enzyme assays.

Spectrophotometric methods: ultraviolet and visible light spectroscopy (principles, instrumentation, applications). Spectrophotometric analysis of enzyme activity: lactate dehydrogenase, creatine kinase, alkaline phosphatase, amino transferase, acetylcholinesterase. Spectrophotometric analysis of metabolites: glucose and cholesterol.

Spectrofluorimetry: principles, instrumentation, applications. Fluorescent probes.

Radioisotopic techniques: the nature of radioactivity, interaction with matter, detection and measurement, instrumentation, safety aspects, application of radioisotopes in the biological sciences.

Electrophoretic techniques: general principles, instrumentation, support media, electrophoresis of  plasma proteins.

Immunochemical techniques: antibodies and their production, antisera, immunoprecipitation and immunodiffusion, RIA, IRMA. Labelling with enzymes: EMIT, ELISA, PEIA. Pregnancy test.

Basic techniques in molecular biology: manipulation of nucleic acids, the polymerase chain reaction, nucleic acid blotting methods, protein blotting methods. Gene expression analysis: the DNA microarray technique. Large scale protein expression alnalysis: the proteomic approach.

Readings/Bibliography

K. WILSON, J. WALKER. Principles and techniques of practical biochemistry. Cambridge University Press, 2000

Teaching methods

During the course tha major biochemical techniques will be discussed, in both theoretical and practical aspects. Examples and problems will be discussed with the students. A practical class (1 CFU) will deal with laboratory experiments and clinical applications.

Assessment methods

Oral examination, to evaluate the theoretical knowledge about biochemical methodologies and the possibility to apply them to clinical analysis.

Teaching tools

PC, overhead projector, biological laboratory

Office hours

See the website of Silvana Hrelia