Protein–ligand interactions are at the core of molecular recognition and have a fundamental role in many scientific areas. Additionally, they are responsible for the main mechanisms of action of many drugs, being important to the tissue distribution, metabolism, and excretion of drugs and bioactive compounds. The aim of this course is the study of protein-ligand binding assays, where ligands range from small organic and inorganic molecules to lipids, nucleic acids, peptides, and proteins. Quantitative analysis of protein-ligand interactions is a fundamental first step towards understanding ligands biological roles. Different ligand-binding assays such as labelled, label-free, thermodynamic and structure-based will be explored. The course will be composed by complementary theoretical and hands-on components, where the limitations of the different methodologies will be discussed and bridged, in order to recognize and overcome incorrect or ambiguous results. The validation of bioanalytical methods will be also explored. The guidelines on bioanalytical method validation (for example the proposed by the European medicines agency) will be focused. Furthermore, experimental protocols will be elaborated and standard operating procedures (SOPs) created.
Learning outcomes
June 12
9h30 - 10h00 - Welcome Reception
Salette Reis - Associate Professor - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
10h00 - 11h00 - Methods to assess protein-ligand binding: theory
Teresa Santos Silva - Assistant Professor - UCIBIO, REQUIMTE, Faculty of Sciences and Technology, Universidade Nova de Lisboa
11h30 - 12h30 - Methods to assess protein-ligand binding: theory
Cláudia Nunes - FCT Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
Catarina Pereira Leite - Post-Doctoral Researcher - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
14h30 - 18h00 - Hands-on training: fluorescence spectroscopy and isothermal titration calorimetry
Cláudia Nunes - FCT Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
Catarina Pereira Leite - Post-Doctoral Researcher - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
June 13
10h00 - 11h00 - Bioanalytical methods validation
Lúcia Saraiva - Auxiliar Professor - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
11h30 - 12h30 - Ligand binding immunoassays: theory
Sofia Costa Lima - FCT Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
Tânia Moniz - Post-Doctoral Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
14h30 - 18h00 - Hands-on training: Ligand binding immunoassays
Sofia Costa Lima - FCT Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
Tânia Moniz - Post-Doctoral Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
June 14
10h00 - 11h00 - Standard operating procedures (SOPs): theory
Sofia Costa Lima - FCT Investigator - LAQV, REQUIMTE, Faculty of Pharmacy, University of Porto
11h30 - 12h30 - Standard operating procedures (SOPs): practice
15h00 - 18h00 - Integrated discussion of SOPs and experimental results