CELLULAR AND BIOCHEMICAL TECHNOLOGIES
Academic year and teacher
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- Versione italiana
- Academic year
- 2022/2023
- Teacher
- MONICA BORGATTI
- Credits
- 6
- Curriculum
- BIOTECNOLOGIE FARMACEUTICHE
- Didactic period
- Secondo Semestre
- SSD
- BIO/10
Training objectives
- The objectives of the course is the acquisition of theoretical principles and their application fields for the main biochemical and cellular methodologies applied in particular to the study of proteins and molecular and cellular interactions.
The acquired knowledge include: the methods used to produce and purify recombinant proteins; techniques used to analyze the amino acid composition, primary structure, concentration, interactions with other biomolecules (nucleic acids, antibodies, transcription factors, etc.) of recombinant proteins; the different cell cultures, how to grow and observe the cells; the embryonic stem cells and iPSCs; the radioisotopes.
The skills acquired through this theoretical course include the knowledge of: the host organisms useful for the production of recombinant proteins used in therapeutic and research applications; the different methods to purify and characterize the proteins according to their chemical-physical and biological properties; the different cellular models used in applied and basic research. Prerequisites
- Basic Knowledges of chemistry, biochemistry and molecular biology are required.
No prerequisites. Course programme
- The course consist of 48 hours of teaching based on frontal lectures.
The course will address the following issues:
- Methods for the production of recombinant proteins: cloning strategies of coding sequences, systems for the expression of recombinant proteins, fusion proteins (6 hours).
-Methods for protein purification (8 hours): separation techniques (extraction of proteins from biological sources; methods of fractionation of protein mixtures; chromatographic techniques) and applications in biomedicine.
- Methods for protein analysis (16 hours): electrophoresis (cellulose acetate, polyacrylamide gel: SDS-PAGE; EMSA assay with and without radiosotopes; footprinting; two-dimensional gels; capillary electrophoresis), quantitative assays (spectrometric and colorimetric methods) , immunochemical techniques (polyclonal and monoclonal antibodies; radioimmunoassay (RIA); ELISA test and applications;
ELISPOT assay and applications; Luminex technology; Western blotting).
-Biochemical technologies for the quantification and analysis of molecular interactions (biosensors) (2 hours).
- Basic principles for working with cells and tissues (16 hours): isolation of cells, visualization of cells and molecules, immortalized and transformed cultures, primary cultures, storage and maintenance of cell cultures, treatments of cells for drug tests, flow cytometry and cell cycle and apoptosis analysis, single cell analysis, blood count with leukocyte formula, embryonic stem cells and iPSCs, cell biobanks.
The hours reported for each topic are indicative. Didactic methods
- The course consists of frontal lectures (48 hours, 6 ECTS), weekly in the classroom using slides in power point format and blackboard provided in the classrooms to explain better the topics.
Learning assessment procedures
- The written exam will be based on multiple choice tests with closed answers consisting of 17 questions by two points for a duration of 25 minutes.
The exam is passed with a score of 18/30.
For DSA students, the test procedure will be evaluated individually. Reference texts
- -Metodologie Biochimiche, Maria Carmela Bonaccorsi, Roberto Contestabile, Martino Di Salvo. Casa Editrice Ambrosiana. Distribuzione esclusiva Zanichelli (2012).
-Biochimica Applicata, ¿Monica Stoppini, Vittorio Bellotti. Edises (2012). ¿¿¿¿¿¿¿¿¿¿¿
-Metodologie di base per le scienze biomolecolari, Rob Reed, David Holmes, Jonathan Weyers, Allan Jones. Zanichelli Editore (2002).
-Biotecnologie molecolari, Terry A. Brown. Zanichelli Editore (2007).
-Biologia molecolare della cellula, Bruce Alberts, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, Peter Walter, Zanichelli Editore (2009).
