Learning outcomes of the course unit
The course aims at enabling the student to understand the key biochemical techniques and to apply the acquired knowledge to the understanding of a biochemical-clinical report. In addition, the basic biochemical knowledge will be applied to the study of the main biological targets of drugs, thus allowing the student to know and understand the molecular basis of drug action.
At the end of the course, the student, also using the previously acquired knowledge of biology, organic chemistry and biochemistry, will need to demonstrate knowledge and understanding about:
- The determination of the main physical and structural properties of proteins, such as the molecular weight, the content of secondary structure, the three-dimensional structure, the isoelectric point, the concentration and the properties of interaction with other macromolecules.
- The application of the basic techniques of molecular biology to the production of recombinant proteins and to site-directed mutagenesis of proteins.
- The main techniques of protein purification from complex samples
- The information contained in clinical biochemistry reports
- The biochemical nature of drug targets
Knowledge of the following disciplines: biology, organic chemistry and biochemistry
Course contents summary
The first part of the course will deal with the main biochemical techniques, mainly those for the expression and purification of recombinant proteins and for protein characterization. In this part of the course, some basic molecular biology techniques will also be covered, with a focus on fundamental techniques for gene cloning and site-directed mutagenesis. The application of biochemical techniques to clinical biochemistry will be discussed at the end of part I.
The second part of the course will be focused on the evaluation of key biological macromolecules and the main metabolic pathways as drug targets, using examples and original literature.
PART I: RECOMBINANT DNA AND ITS APPLICATIONS
• Recombinant DNA technology
• Expression of recombinant proteins
PART II: PROTEIN PURIFICATION AND ELECTROPHORESIS
• Purification of proteins
PART III: CHARACTERIZATION AND IDENTIFICATION OF PROTEINS
• Spectroscopic techniques
• Enzyme kinetics at the steady-state
• Immunochemical techniques
• DNA sequencing
• Introduction to proteomics
PART IV: a brief introduction to clinical biochemistry
Clinical biochemistry: definition.
The biochemical and clinical examination: The biological samples. The analytes. The measured values. The units of measure. The reference values.
Clinical biochemistry in major metabolic disorders and major organ dysfunction (renal, cardiac, liver and blood).
Biochemistry of drug targets
The nature of drug targets
DNA as a drug target: Drugs that interact with DNA.
RNA as a drug target: antisense nucleotides and structural analogues.
Drugs that interact with ribosomal RNA and other ribozymes.
Membrane lipids as drug targets.
Proteins as drug targets: Enzymes; receptors and signal transduction pathways, transmembrane carriers; pumps and channels
Course material provided by the teachers.
- Biochimica e Biologia Molecolare - Principi e tecniche. A cura di Keith Wilson e John Walker. Raffaello Cortina Editore.
- Metodologie Biochimiche. A cura di Carmela Bonaccorsi di Patti, Roberto Contestabile e Martino Luigi di Salvo. Casa Editirce Ambrosiana.
- “Stryer – Biochimica”. A cura di Berg, Tymoczko. Zanichelli editore
The course will deal with the principal biochemical and molecular biology techniques, also applied to clinical biochemistry. Due to the peculiar nature of the course, the teacher, when possible, will integrate the theoretical lessons with short demonstrations, also with the support of short videos. Experts in cutting-edge techniques will be invited to give seminars.
In the second part, lectures will be integrated with one-to-one discussions with the teacher about original articles of the scientific literature.
Assessment methods and criteria
The assessment of the achievement of the objectives will include an oral examination divided into two parts. The first part will include questions relating to the main biochemical techniques and will establish whether the student has attained sufficient knowledge of these techniques, even in relation to the ability to apply that knowledge to the solution of practical problems. The second part will cover the biochemistry of drug targets, with the aim of stimulating the student to understand the molecular mechanisms that underlie the action of drugs. Students may be required to read and discuss scientific papers provided by the teachers.
At the end of the examination two separate evaluations will be given, whose average will be the final grade.
In order to pass the exam, the student must demonstrate to be able to autonomously discuss, using appropriate technical language, all the topics covered in the course, at least at a basic level. Higher levels of detail, understanding of the examples and the careful study of additional material will lead to a higher evaluation.