8h30 – 8h50
Credenciamento
8h50 – 9h
Abertura
Ana Carolina M Figueira – LNBio/CNPEM
9h-11h
Microcalorimetry: A versatile tool for the characterization of biomolecular interactions – Dr Raúl Pacheco-Gómez. Application Scientist – Malvern Instruments, UK
Higher-order biomolecular structures and their dynamic interactions with various ligands and biomolecules drive and regulate all biological processes; studies of biomolecular interactions are fundamentally important in all areas of life sciences. A rigorous understanding of any biological interaction requires quantification of its thermodynamic and kinetic properties. This, combined with structural detail, can be assimilated to enable a picture of the change (for example, going from the free to the bound state in a biomolecular interaction, or from the folded to the unfolded state of a biomolecule). Isothermal Titration Calorimetry (ITC) and Differential Scanning Calorimetry (DSC) provide the thermodynamic data required for developing this understanding. ITC is the ideal technique for the measurement of biological binding interactions since the data provided does not rely on the presence of chromophores or fluorophores, nor requires an enzymatic assay; ITC relies only on the detection of a heat effect upon binding. ITC and DSC are label-free, enabling scientists in academia and industry to better understand the conformational stability of their biomolecules and their binding to biologically relevant interactants. This presentation covers the principles of ITC and DSC and it exemplifies a broad range of applications enabled by the direct nature of the technique. Experimental practicalities as well as how to obtain quality data will be covered.
11h-11h30h Coffee Break
11h30-12h30 – Pensabio e suas soluções para caracterização de proteínas – Dra Danuza Rossi.
12h30 – 14h00 – Almoço livre
14h00- 15h00 – Data treatment and software presentation – Dr Raúl Pacheco-Gómez. Application Scientist – Malvern Instruments, UK
A special focus will be given to the benefits of PEAQ-ITC and PEAQ-DSC, the latest generation of MicroCal instruments, and the solutions they offer for addressing current bottlenecks associated with the study of interaction analysis and conformational stability. Among the most recognized challenges is the need to adequately address a broad range of binding affinities and to reliably interpret the binding data, complicated by the presence of inactive protein or inherent uncertainty in the concentration of the ligand. We will discuss the improvements in PEAQ-ITC and PEAQ-DSC data quality which enable increased confidence and data resolution when measuring low heats at low or uncertain sample concentrations and complex binding modes. The new MicroCal PEAQ-ITC and PEAQ-DSC analysis softwares will also be presented: The new softwares allow for automated data analysis, minimizing analysis time and user subjectivity in assessing data quality as well as allowing the analysis of large data sets of 50 or more experiments in a matter of seconds.