Protein Engineering and Proteomics
A.Y. 2021/2022
Learning objectives
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.
Lesson period: Second semester
Assessment methods: Esame
Assessment result: voto verbalizzato in trentesimi
Single course
This course cannot be attended as a single course. Please check our list of single courses to find the ones available for enrolment.
Course syllabus and organization
Single session
Responsible
Lesson period
Second semester
Teaching methods: The lessons will be held on the Microsoft Teams platform and can be followed both synchronously on the basis of the second semester timetable and asynchronously because they will be recorded and left available to students on the same platform. The schedule of lessons and all the details of the activities will be published through notices on the ARIEL board. before the start of the lessons. Any updates will also be communicated through notices on the ARIEL board.
The methods and criteria for participating to any possible in classroom lessons and practical labs as established by the University, will be published in due time on the ARIEL notice board of the course. The exercises will take place partly in person and partly online, both asynchronously (providing the material with audio commentary through the Ariel site) and with synchronous mode to critically discuss publications and scientific reports of interest (through the Microsoft Teams platform ).
Program and reference material: The program and reference material will not change.
Learning verification procedures and assessment criteria: The exam will take place orally using the Microsoft Teams platform, according to the indications provided by the University.
More specific information on the delivery mode of training activities for academic year 2021/2022 will be provided over the coming months, based on the evolution of the public health situation.
The methods and criteria for participating to any possible in classroom lessons and practical labs as established by the University, will be published in due time on the ARIEL notice board of the course. The exercises will take place partly in person and partly online, both asynchronously (providing the material with audio commentary through the Ariel site) and with synchronous mode to critically discuss publications and scientific reports of interest (through the Microsoft Teams platform ).
Program and reference material: The program and reference material will not change.
Learning verification procedures and assessment criteria: The exam will take place orally using the Microsoft Teams platform, according to the indications provided by the University.
More specific information on the delivery mode of training activities for academic year 2021/2022 will be provided over the coming months, based on the evolution of the public health situation.
Course syllabus
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 protected].
Teaching methods
Frontal lessons, laboratories, numerical exercises in classroom and simulations in computer room.
Lessons: 36 hours (4.5 CFU)
Practical labs: 8 hours (0.5 CFU)
Exercises: 16 hours: (1 CFU)
Lessons: 36 hours (4.5 CFU)
Practical labs: 8 hours (0.5 CFU)
Exercises: 16 hours: (1 CFU)
Teaching Resources
Slides of the lessons will be available to students through ARIEL platform. https://ascarafonipep.ariel.ctu.unimi.it/v5/home/Default.aspx
Suggested textbooks:
"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
Suggested textbooks:
"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
Oral. At least 7 exam sessions per year will be guaranteed. The date of the exam will be published on SIFA platform.
Oral 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
Specific procedures for students with disabilities or specific learning disabilities (DSA) will be applied also for telematic exams. Here the complete information:
https://www.unimi.it/en/study/student-services/services-students-disabilities
https://www.unimi.it/en/study/student-services/services-students-specific-learning-disabilities-sld
In case you need specific procedures, please inform the teacher by mail at least 10 days before the exam, including in the addresses [email protected] or [email protected].
Oral 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
Specific procedures for students with disabilities or specific learning disabilities (DSA) will be applied also for telematic exams. Here the complete information:
https://www.unimi.it/en/study/student-services/services-students-disabilities
https://www.unimi.it/en/study/student-services/services-students-specific-learning-disabilities-sld
In case you need specific procedures, please inform the teacher by mail at least 10 days before the exam, including in the addresses [email protected] or [email protected].
BIO/10 - BIOCHEMISTRY - University credits: 6
Practicals: 8 hours
Laboratories: 16 hours
Lessons: 36 hours
Laboratories: 16 hours
Lessons: 36 hours
Professor:
Scarafoni Alessio
Professor(s)
Reception:
Thursday afternoon only by appointment (by e-mail)
Building 21040, first floor