Protein engineering, drug delivery and regulatory aspects

A.Y. 2019/2020
Overall hours
Learning objectives
The course aims to explore the development, formulation, manufacturing and marketing of Biotechnological and Advanced Therapy Medicinal Products. The course is divided in three modules covering: (i) the key-concepts and tools to understand the molecular strategies behind the rational design of proteins having new functions or activities, (ii) the biopharmaceutic aspects and the design of drug delivery systems intended for the main routes of administration, and (iii) the regulatory aspects related to their manufacturing and marketing.
Expected learning outcomes
At the end of the course, students will have acquired the knowledge and methods enabling them to study and think, critically and independently, about different issues of medicines for human use, with reference to biotechnological medicinal products. The students will learn how to rationally design, or modify, protein structures and activity. They will know the basis on how to approach this problem, and to propose solutions for implementing and improving protein functions according to a specifically requested need. They will know how to edit the genome of cells and even organisms with the aim of potentially crate new GMOs. Also, they will know how to set-up experiments for the high-scale production of proteins and characterize them post-production.
Based on the physico-chemical features of an active ingredient and the selected parenteral route of administration, the student will be able to choose the optimal technological approach to the design and manufacture pharmaceutical dose forms satisfying the criteria of quality, safety and efficacy.
Moreover, students will be able to recognise what is, and what is not, a medicinal product, and outline the steps required to place a medicinal product on the market, with particular reference to biotechnological and advanced therapy medicinal products. They are expected to be able to retrieve and understand the relevant parts of EU legislative acts.
Students will be also able to correctly understand the questions in the learning verification phase, to know how to elaborate and connect the knowledge acquired in all the didactics units of the course.
Course syllabus and organization

Single session

Lesson period
First semester
Course syllabus
The course on "Biopharmaceutics and drug delivery" aims to examine the basic principles of the main routes of administration and to discuss their importance in designing a medicinal product.
· In-depth analysis of pharmacokinetics and biopharmaceutical aspects of the routes of administration.
· Parenteral dosage forms: formulative approaches and quality controls.
· Stability of a medicinal products.
· Water for pharmaceutical purposes: classification, requirements, production methods.
· In-depth analysis of sterilization methods: filtration, heat sterilization, irradiation
· Lyophilization
· Clean rooms and aseptic preparation.
· GMP (Annex 1 and 2).
The lessons on "Regulatory aspects" focuses on the regulatory issues related to the manufacturing and marketing of medicinal products.
EU Institutions and the European Medicines Agency (EMA).
Definition of medicinal product. Borderline products
EU Pharmaceutical Law.
Manufacturing Authorisation and Marketing Authorisation. Biosimilars.
The Common Technical Document.
Advanced Therapy Medicinal Products (ATMPs).
ATMPs regulatory aspects.
Orphan medicines and drug repurposing. Medicines for children.
Labelling of medicinal products. Pharmacovigilance.
Market access.

The program on Protein Engineering is characterized by 3 distinct sections. The first will revisit some basic concepts of biochemistry and molecular biology with a focus on the engineering aspect, and will include: protein nature and structure, transcription, protein production, protein maturation and modifications, plasmid production and expression, protein activity, restriction enzyme and cloning libraries. The second section will introduce some of the most used methodologies and techniques in protein engineering and will include: PCR, qPCR, direct evolution, random mutagenesis and error prone PCR, gene editing (CRISPR/Cas9 and TALEN technology), phage display for protein and ligand discovery, principles of optogenetics. The last section will list the most important protein characterization techniques like: 2D gel electrophoresis (also in DIGE modality), mass spectrometry, Western Blot and immunoprecipitation techniques, immunofluorescence, x-ray crystallography, NMR, cryo-TEM, liquid TEM.
Prerequisites for admission
Prospective students are required to be familiar with the basic concepts of inorganic and organic chemistry, biochemistry and molecular biology as well as general knowledge on physiology. These topics are generally provided within courses at the Bachelor Degree.
Teaching methods
Lectures and practical classes in laboratory
Teaching Resources
Lecture handouts will be available on the ARIEL website.
Aulton's Pharmaceutics, The Design and Manufacture of Medicines
Edited by Michael E. Aulton and Kevin M.G. Taylor
Relevant guidelines for the Regulatory aspects are listed on the ARIEL website (in the 'Course information' section).
For the protein engineering, the students are encouraged to read Lewin's GENES XII 12th Edition by J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick (Author) (earlier editions are also suitable) and Molecular Biotechnology: Principles and Applications of Recombinant DNA, Fifth Edition by B.R. Glick and J.J. Pasternak
Assessment methods and Criteria
The exam will consist in both an oral and a written test. The students will be evaluated based on their ability to answer theoretical questions about the subjects taught during the course and to frame them in a wider scenario. Moreover, students will be evaluated based on their ability to establish connections among the different subjects dealt with during the course. The oral test will be focused on questions related to "Biopharmaceutics and drug delivery" and "Regulatory aspects"; th students should show their capability to clearly describe the design of drug delivery intended for parenteral administration: from the selection of excipients to the process development in order to satisfy all requirements, to the regulatory aspects of manufacturing, marketing and post-marketing surveillance. The oral test will contribute to 65% of the final grade.
Regarding Protein Engineering, the exam will be based on a written examination only. Here the students will be asked 2 theoretical questions on basic processes of protein engineering and 1 question will include a methodological procedure and/or protein characterization techniques. The exam will last for 3 hours. Any possible changes in the examination procedure (in terms of number and type of question) will be discussed during the lectures of protein engineering and agreed with the students.
The exam can be finalized only if both the oral and written tests are passed, and the final grade will be the normalized average of the two grades.
Biopharmaceutical and formulation aspects
Single bench laboratory practical: 16 hours
Lessons: 24 hours
Professor: Selmin Francesca
Chemical biology and protein engineering
Lessons: 32 hours
Professor: Rizzello Loris
Regulatory aspects of medicinal products
Lessons: 24 hours
Professor: Rocco Paolo
by appointment.
Dept. Pharmaceutica Sciences - via G. Colombo, 71