The course provides an introduction to protein biotechnology and the basic knowledge required for set up large-scale protein production within biotechnological industry, from cell cultivation in bioreactors up to protein purification in a preparative scale and includes different modern bioanalytical techniques. The course is aimed at giving students in-depth understanding of 1) the structure-function relationship of proteins and of 2) the methodologies used to produce, characterize and use proteins, enzymes included, and 3) the molecular basis and protein engineering tools available for designing enzymes or proteins with new or desirable functions. The course will have a strong hands-on connotation based on practical sessions either of experimental design in classroom and of bioinformatics.
Expected learning outcomes
At the end of the course the student will be able to implement the notions of molecular biology, proteomics and genetic engineering for the production and improvement of proteins and enzymes. The student will be able to analyze the structure of proteins and their post-translational modifications; review the main factors that are significant for protein folding processes and stability; explain how proteins can be used for different industrial applications, carry out mutagenesis approaches to improve protein stability and to confer on them new functions.
Structural/functional molecular characteristics of proteins: from biosynthesis to folding. Structural and functional dynamics of proteins. Modern approaches for the study of the proteins. Implementation of protein purification procedures. Proteome analyses for protein and enzyme identification and exploitation. Recombinant protein technologies; gene expression: vectors, expression in bacteria, yeasts, plants, insect and mammalian cells. Cell-Free translation systems. Inclusion bodies, their solubilization; protein refolding. Co-expression and use of chaperones. Storage stabilization of proteins. Hints of evolutionary biotechnology (rational design and directed evolution). Examples of tailored enzymes in industrial biocatalysis. Production and applications of polyclonal and monoclonal antibodies. Strategies to set up a protein engineering project. Proteomics: principles and methodologies.
Prerequisites for admission
Prerequisites: general knowledge of structural biochemistry (protein primary, secondary, tertiary quaternary structures, including basics of protein folding) and cell protein synthesis. Enzymatic activities. Basics of gene cloning and heterologous gene expression. For any question and further details please contact the teacher at email@example.com.
Frontal lessons (3.5 CFU), laboratories (1.5 CFU), numerical exercises in classroom and simulations in computer room (1.0 CFU).
Slides of the lessons will be available to students through ARIEL platform. "Protein Engineering" by Moody P C E and Wilkinson A J, "Proteins" by Creighton T E, "Introduction to Protein Structure" by Branden C and Tooze J., "Biochemistry" by Voet D and Voet G, "Protein Engineering" by Kurra Venkata Gopaiah, "Medicinal Protein Engineering" by Yury E Khudyakov, "Protein Engineering: Design, Selection and Applications (Protein Biochemistry, Synthesis, Structure" by Mallorie N Sheehan, "Protein Engineering: Principles and Practice" by Jeffrey L Cleland and Charles S Craik "Protein Engineering Handbook", 1 & Volume 2, by Lutz and Bornscheuer
Assessment methods and Criteria
Written. At least 7 exam sessions per year will be guaranteed. The date of the exam will be published on SIFA platform. Written exam based on questions about the topics covered during the course. The learning verification will be evaluated according to the following criteria: 1. Proof of understanding of the topics covered during lectures and laboratory practices 2. Ability to expose, in a critical and integrated way, the molecular determinant of proteins structure and their post-translational modifications; the main factors that are significant for protein folding processes and stability; how proteins can be used for different industrial applications; how to carry out mutagenesis interventions to improve protein stability and to confer on them new functions. 3. Knowledge of laboratory methodologies to assess protein structure, functionality and stability. 4. Correct terminology 5. Completeness of the answers