APPLIED BIOCHEMISTRY
Academic year and teacher
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- Versione italiana
- Academic year
- 2015/2016
- Teacher
- MARCELLO BARONI
- Credits
- 6
- Curriculum
- Biologia molecolare e cellulare
- Didactic period
- Primo Semestre
- SSD
- BIO/10
Training objectives
- The study of the theoretical basis, instrumental implementations and major experimental approaches in biomedical sciences; preparation and analysis of samples for biochemical studies.
Knowledge and understanding.
The student:
- knows physical and chemical principles at the base of the main techniques and instruments used in biochemical investigations;
- knows the parameters that can act to modify a biochemical investigation;
- includes protocols and ways to optimize them;
- is able to describe the structural elements of the main instruments of a biochemical laboratory;
- knows the terminology used in biochemical methodologies.
Ability to apply knowledge and understanding.
The student:
- is able to use the knowledge gained to evaluate the results of an experiment, identify issues and optimize it;
- assess the potential impact of changes in parameters involved in biochemical experiments and major clinical investigations;
- is able to discriminate against qualitative and quantitative methodologies, selecting the appropriate.
- knows how to properly use the terminology of the biochemical methodologies. Prerequisites
- There are no prerequisites.
A solid background of Biochemistry, General Chemistry, Laboratory of Chemistry, bases of Physic, Molecular Biology and of Immunology are recommended, thus achieving a complete understanding of the proposed program. Course programme
- IN THE FIRST PART OF THE THEORETICAL COURSE (about 20 hours):
general principles, instrumentations and applications of the major separation and analytical experimental approaches:
Electrophoretic techniques (SDS-PAGE, Native-PAGE, IEF, 2D, Capillary).
Protein and nucleic acid purifications.
Chromatographic techniques (ion exchange, gel filtration, affinity, immunoaffinity, absorption). Centrifugative techniques.
Principles, instrumentation and applications of spectrophotometric techniques (ultraviolet, visible and infrared light) and Spectrofluorimetry. Fluorescent probes and substrates for enzymatic kinetics.
Protein structural and functional studies.
Immunochemical principles and techniques (Immunoprecipitation and immunodiffusion, SRID, Rocket, Western Blotting, E.L.I.S.A.). Antibodies and their features.
Radiochemical techniques (R.I.A.).
Signal amplification systems based on Tyramide molecule.
Sample preparation for 2D analysis.
IN THE SECOND PART OF THE THEORETICAL COURSE (about 16 hours):
- Cell culture studies.
- Fluorescence microscopy
- Cytofluorimetry and ImageStream
- Xcelligence, a Realtime Cell Analyzer
IN THE THIRD PART OF THE THEORETICAL COURSE (about 4 hours) they will be considered the innovative techniques, based on microspheres:
- purifications with Dynabeads.
- Simultaneous multiple dosages by Bioplex.
- Protein equalization by Proteominer.
- Interaction studies with amplified luminescent proximity homogeneous assay (alpha assays). Didactic methods
- The major biochemical techniques will be discussed, both in theoretical and in practical aspects. The ex cathedra teaching will made with slide support, projector and biological laboratory.
Examples and biochemical problems will be discussed with the students. 1 ECTS (12 h) of a practical class will deal with laboratory experiments (E.L.I.S.A., Western Blotting, Protein staining, Spectrophotometry) to make the student familiar with the laboratory equipments and problems. The course is designed to understand the technology and scope of measuring techniques in biochemistry and apply them to industrial productivity and research, with accuracy and precision. The students are also encouraged to develop their critical thinking and communication skills through stimulation of active participation to the lesson. Learning assessment procedures
- Evaluation at the end of the semester will consist of one final and written test, with five or six open questions (5 or 6 points), covering the theoretical knowledge of the main subjects of the course, about biochemical methodologies and the possibility to apply them to clinical and research analysis.
The examination, using both calculations and short answer questions, will allow students to demonstrate their proficient and flexible knowledge in the biochemistry methodologies.
The exam will take between 50 and 60 minutes. Reference texts
- - Biochimica Applicata - Monica Stoppini e Vittorio Bellotti - Edises.
- Metodologie Biochimiche - M.C. Bonaccorsi di Patti, R. Contestabile, M.G. Di Salvo - Ambrosiana.
- Principi di metodologia Biochimica - Carlo De Marco e Chiara Cini - Piccin.
- Biochimica e Biologia Molecolare. Principi e tecniche Wilson K, Walker J. Zanichelli.
- The Protein protocols Handbook - Walker, John M.-
Biochimica - J. M. Berg, J. L. Tymoczko e L. Stryer. Zanichelli.
- Membrane Protein Protocols - Selinsky BS.
- Metodologie di base per le scienze biomolecolari - Reed R, Holmes D, Weyers J, Jones A. Zanichelli.
Scheme and Figures provided.
