The course aims at exposing PhD students with a background in pharmaceutical sciences to the emerging field of biopharmaceuticals, their production, characterization and derivatization. The students should be able to read critically a research paper detailing the properties of a biopharmaceutical and to understand the most common problems associated with their production, particularly in comparison with small-molecule drugs.
Course contents summary
The course will offer an overview of proteins from a biotechnological point of view, including their production, purification, modification, and characterization (through enzyme assays, fluorescence, circular dichroism spectroscopy).
1) Protein expression in bacteria, plants, insect and mammalian cells
The main recombinant expression systems will be discussed, with particular emphasis on those currently yielding commercially available products and on novel and future prospects. The laboratory and industrial fermentation systems will be presented.
2) Protein extraction and purification
Affinity tagging of recombinant proteins and the corresponding separation methods will be presented. State-of-the-art chromatographic techniques will be discussed.
3) Protein chemistry, bioconjugates and protein formulation
Protein chemical modification, protein PEGylation, reactive sites, site-specific modification, chemical modification effects on protein structure, function and stability will be discussed.
4) Protein characterization using biochemical and biophysical methods with special emphasis on enzyme kinetics
Enzyme kinetics, determination of catalytic parameters, enzyme inhibition and determination of inhibition mechanism and IC50/Ki, assay considerations for compound library screening; analysis of secondary, tertiary and quaternary structure and protein thermodynamic stability by circular dichroism spectroscopy; fluorescence, fluorescence quenching and FRET as a tool to investigate protein-protein and protein-ligand interactions, protein conformation and dynamics, protein microenvironment.
Material provided by the teachers. A few representative open access scientific papers/reviews focusing on the main topics of the course will be indicated.
In the in-person lectures, teachers will illustrate and discuss the basic principles and the theoretical aspects of the different topics of the program. Students will then visit the laboratories of the teachers to be introduced to the instrumentation and receive practical demonstrations of the relative experimental techniques. If restrictions to the accessibility of laboratories will still be in place, remote demonstrations or alternative activities will be organized.
Assessment methods and criteria
At the end of the course, students will have to report on theoretical and practical aspects of some of the experimental techniques illustrated by teachers, also trough critical reading and discussion of selected scientific papers.