O Workshop
O Laboratório Nacional de Biociências (LNBio), em parceria com a Malvern e a Pensabio, promove o Workshop em Calorimetria de Proteínas e Técnicas Ortogonais para Caracterização de Biomoléculas. O evento será realizado em 22 de junho, no campus do Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), em Campinas-SP. A iniciativa é gratuita e as inscrições devem ser feitas até 16 de junho.
O workshop discutirá os dois principais tipos de calorimetria de proteínas – calorimetria de titulação isotérmica e calorimetria de varredura diferencial. A programação do evento abordará os mais recentes avanços destas tecnologias e apresentará como estes métodos biofísicos são utilizados nas pesquisas das áreas de saúde e ciências biológicas. As aplicações da calorimetria no processo de descoberta e desenvolvimento de fármacos baseados em pequenas moléculas e biofármacos também serão tema de discussões. A ementa do Workshop compreenderá ainda técnicas auxiliares para a determinação de peso molecular, viscosidade e concentração, que contribuem para a caracterização de fórmulas proteicas voltadas à terapêutica.
Program
9:00 – 10:45
Introduction of Dr. Ronan O’Brien
Ronan O’Brien is Head of Business Development for the MicroCal products at Malvern. He has a PhD in biophysical chemistry and spent many years in the area of label free interaction analysis, a discipline in which he has published widely and includes contributions to a number of educational textbooks. He carried out his post doctoral research with Professor Julian Sturtevant at Yale University and Professor John Ladbury at University College London before joining MicroCal as an applications expert in 2001. Since then he has held a number of roles including Head of R&D and Scientific Marketing Manager.
Isothermal Titration Calorimetry and Differential Scanning Calorimetry in Life Sciences Research and Drug Discovery and Development: Binding, Thermodynamics and Stability
Essentially all biological processes involve changes in inter- and intramolecular interactions. The forces involved in these interactions are quantitated by thermodynamic parameters, and are part of the complete structure-function-thermodynamics description of biomolecular interactions. These processes involve proteins (including enzymes and antibodies), nucleic acids, lipids, (including biological membranes), polymers, as well as nanoparticles, surfactants, carbohydrates, and other molecules. Thermodynamics are used to characterize interactions as well as an aid in the discovery, design and development of new drugs.
Differential scanning calorimetry (DSC) is used to study intramolecular interactions, such as protein unfolding and stability. Isothermal Titration Calorimetry (ITC) measures the enthalpy change, binding affinity, and stoichiometry of an intermolecular binding reaction, such as a protein-ligand or antibody-antigen interaction. ITC and DSC are label-free methods, using native molecules in solution, and uses heat change as the detection method.
This workshop introduces ITC and DSC, discusses recent advances in the technology, and shows how these biophysical methods are used in life science research, and discovery and development of small molecule drugs and biopharmaceuticals.
10:45- 11:00
COFFEE BREAK
11:00 – 12:00
A new approach to improving early characterization of therapeutic protein formulations
Geisi Rojas Barreto – Product Specialist at Pensabio Biotecnologia
Measurements: Molecular Sizing – TDA with UV absorption; Ligth Scattering; Resonant Mass Measurement; Viscosity – Microcapillary flow and Poiseuille’s Law; Concentration – Nanoparticle Tracking Analysis and Resonant Mass Measurement
Applications: Size and Viscosity– Small molecules; Proteins in the presence of aggregates; Proteins in the presence of excipients.
The influence of formulation components such as excipients and surfactants on protein stability and self-association in solution is important to understand, as biotherapeutic products generally require complex formulations. TDA (Taylor Disperson Technique) is a fast and inherently simple, label-free and solution based technique for sizing small molecules through to protein-based systems. Combined with UV detection, it offers specificity for a target molecule and, by referencing against the sample buffer, is unaffected by the presence of excipients or surfactants. Mass-weighted detection means that the presence of larger species or aggregates does not dominate the measured data, and TDA can therefore track changes of target molecule species in aggregated solutions. Measurement capabilities at low concentrations, and with ultra-low sample volumes, makes TDA a powerful orthogonal technique for studying the association and conformation of proteins in complex formulations and media, with features that are highly complementary to the well-established DLS technique.
Protein aggregation – Bridging the sub micron gap
The formation of protein aggregates is a particular concern for parenteral administration biopharmaceuticals due to increased immunogenicity. As a consequence, there is an expectation from regulatory agencies for companies to monitor levels of subvisible particles present in therapeutic protein products initially and over the course of the shelf life. Increasingly, interest has focused on subvisible particles between 0.1 – 10 µm, a size range that has, up to recently, been challenging to study. The development of Resonant Mass Measurement technology in the Archimedes system provides a unique solution to this challenge, delivering quantitative data for particles between 0.2 – 5 µm as well as the ability to distinguish between negatively buoyant particles, such as protein aggregates, and positively buoyant particles such as silicone oil.
Protein stability using Raman – Combining dynamic light scattering and raman spectroscopy to achieve new insights into protein stability, aggregation and high order structure
The technique uniquely combines two well-established analytical techniques namely dynamic light scattering (DLS) and Raman spectroscopy to derive structural, thermodynamic and kinetic insights into the mechanisms of protein aggregation and the factors that influence protein stability.
12:00 – 1:30
LUNCH TIME
1:30 – 3:00
Communication Sessions: Projects in Development and Challenges From LNBio Researches
3:00 – 4:00
Topics Discussions
4:00 – 4:30
COFFEE BREAK