Ateneo - UPO

The course through a theoretical and practical approach in the laboratory provides a broad focus on the main biochemical and biomolecular methodologies.

The teaching material offered on the Moodle platform of the course is an additional support to the course. Material for any further information will also be suggested during the course, including specific papers describing the techniques described during the course.
Principal text: “Metodologie biochimiche e biomolecolari” Mauro Maccarone Zanichelli

The mission of the Applied Biochemistry course is to provide students with knowledge of the main biochemical/biomolecular methodologies with particular attention to the determination of characteristic parameters in the study of proteins. The student will deepen 1. the theoretical and instrumental biochemical bases of the main analytical and preparative methodologies for the purification and analysis of proteins; 2. the bases of biomolecular and biochemical techniques applied to biomedical research. 3. In the practical part of the course, the student will acquire basic knowledge of the instruments present in a laboratory and the ability to carry out some experimental biochemical assays.

Good knowledge of the principles of Biochemistry and Molecular Biology

Class lessons and laboratory sessions.
Attendance in the laboratory is highly recommended

Discussion of the program's arguments in class. The course includes the execution of sessions in the laboratory. At the end of each experiment we proceed directly with students in the testing and discussion of the results obtained. The course is supported in DIR section (Applied Biochemistry ) with use of the student material for verifying the level of initial and ongoing learning preparation . More details are available on all the experimental methods of the course.

The objective of the exam consists in verifying the level of knowledge and deepening of the topics of the course program and the reasoning skills developed by the student. The student must keep a laboratory notebook to develop the ability to describe a completed experiment and collect data correctly. Student must also produce a final written report containing a critical analysis of the results obtained in the experiences to develop the ability to draw conclusions from the experiences and communication skills. The examination focuses on the priciples of the methods and the discussion of the experiments presented in a laboratory notebook in which the methodological principles and the experimental data will be evaluated. Moreover during the course the students, divided in several groups, will analyze and prepare a presentation on a scientific paper describing an application of the methods discussed in the course.
The evaluation will include the lab book, the relationship and the oral exam and the presentation of the paper and is expressed in thirtieths (minimum mark 18/30). Sufficiency is achieved by proving to know and understand the basics and their applications in the laboratory. Excellence can be achieved by demonstrating strong qualities both in producing a flawless relationship and in an adequate manual and ability to conduct the experiments carried out that will be critically commented on.

During the course the following topics will be illustrated, some deepened with practical exercises/experiments in the laboratory which include the preparation of a laboratory notebook and a final individual report:
1. Equipment and laboratory instrumentation, safety in the chemical - biological laboratory. Quantitative measurements in Biochemistry. The laboratory notebook and the setup of an experiment.
2. Preparation and handling of a biological sample: use and preparation of buffer solutions; cell lysis; principles of centrifugation: centrifuges, rotors, differential and density gradient centrifugation, extraction techniques. Cell cultures: primary and secondary cultures, cell lines. Methods of cell culture.
3. Molecular Biology and techniques (Nucleic Acid Structure, Genes and Genome, Nucleic Acid Function, Nucleic Acid Handling, Basic Tools and Analysis Techniques, Isolation and Separation of Nucleic Acids, Polymerase Chain Reaction (PCR), analysis of the nucleotide sequence, analysis and manipulation of gene expression (recombinant DNA and transfections), analysis of the whole human genome.
4. Protein purification techniques and analysis: structure, purification procedures, characterization and analysis of protein functions, analysis of the ionic properties of amino acids and proteins. Chromatographic techniques, electrophoretic techniques and qualitative and quantitative assays of protein (spectrophotometric methods).
5. Immunochemical techniques. Notions on antigen and antibody, monoclonal antibodies and their preparation. Immunoblotting and immunoassays. Immunoassay systems based on immunoprecipitation and labeling of antigens or antibodies with enzymes (ELISA) and (RIA).
6. Introduction to immunofluorescence, immunocytochemistry, microscopy, cytofluorimetric analysis and main applications.
7. System Biology. Basic principles of mass spectrometry: outline of proteomics and metabolomics and applications. Bioenergetics.

Knowledge and ability to understand. Acquisition of in-depth knowledge on the main techniques applied to biochemistry and molecular biology. Acquisition of the mastery of the theoretical tools for the interpretation of biochemical processes. Acquisition of appropriate scientific language. Ability to apply knowledge and understanding: ability to collect data correctly and to keep a laboratory notebook; ability to apply theoretical knowledge to the execution and understanding of laboratory experiments and to the interpretation of the results obtained. Autonomy of judgment
Acquisition of independent judgment in the evaluation of experimental data identifying possible errors and proposing solutions concerning problems in the biochemical laboratory.
Communication skills
Improvement of the disciplinary vocabulary in the biochemical field, as well as the ability to describe, with clarity and critical sense, of phenomena and biochemical problems even to non-professionals. Ability to acquire information on university texts and research papers in English.
Learning skills
Acquisition of the ability to deepen and independently update the knowledge in the subject, through reading texts and scientific articles. Ability to use the teaching material for a critical and reasoned study.