The third European MicroCal meeting was held on 13th and 14th September in the beautiful venue of the Institute of Engineering & Technology (IET) at Savoy Place, on the banks of the River Thames in London. We welcomed delegates from across Europe to share and discuss their calorimetry expertise in an agenda packed with excellent scientific presentations.
The meeting kicked off with a morning workshop which was repeated in the afternoon session. The workshop’s focus was firmly on data quality and the importance of establishing sample integrity prior to further characterization. This was emphasized by Stephan Uebel, who runs a core facility at the Max Planck Institute of Biochemistry. Stephan highlighted the importance of establishing sample quality, concentration and purity – with no aggregates present – prior to any other biophysical characterization such as Isothermal Titration Calorimetry (ITC). Natalia Markova, Principal Scientist for MicroCal, added her expertise to this session, prompting lots of discussion from the delegates.
Hanna Jankevics Jones, Principal Applications Scientist at Malvern Panalytical, hosted the next session and highlighted the need to check and confirm protein stability, ensuring sample integrity. Hanna discussed Dynamic Light Scattering (DLS) and how the information this technique provides, such as the second virial coefficient (B2) and zeta potential, can be used to predict colloidal stability. Showing a case study of the use of DLS and Size Exclusion Chromatography (SEC) to assess the stability of recombinant human serum albumin samples, Hanna highlighted the factors that need to be considered to fully understand colloidal stability and thermal stress. Hanna also introduced the new Zetasizer Ultra, with its new functionality to assist in assessing sample integrity prior to further characterization.
Margarida Bastos, of Porto University, and Natalia Markova jointly led the next session, which was focused primarily on Differential Scanning Calorimetry (DSC), a first principal technique for measuring functional stability, and ‘Stability beyond TM‘. Two case studies were presented in this session, showing how parameters other than TM can be used to assess thermal stability. For example, Tonset is an additional parameter which provides valuable information for assessing thermal stability and aggregation propensity. Margarida also outlined how DSC is an excellent tool for characterization of lipids and liposomes.
Peter Gimeson, Bioscience Advocacy Manager at Malvern Panalytical, led the next session in discussion of ITC and how sample preparation/quality is the key to successful ITC experiments. Examples were shared showing how ITC has been used to measure DNA–protein binding. The investigation of enzyme kinetics using ITC was explored and compared to data obtained with conventional methods. Tips and tricks concerning how to optimize ITC experiments were shared amongst the audience.
This was followed by a session led by Adrian Velazquez- Campoy (Zaragoza University), Niek Buurma (Cardiff University) and Eva Muñoz (Santiago de Compostela University), in which complex ITC data and quality assurance were discussed. Basic concepts were followed by more in-depth discussion about the challenge of multiple interaction sites and the complexity of fitting data. The importance of validating the fitting of data was discussed in much detail, with great engagement from the audience.
The workshop was rounded off with the ‘Ask the Expert‘ session, where the panel answered questions from the audience. The session was very interactive with lots of debate on how sample quality is important and how to understand if aggregates are affecting the sample integrity. Concentration and purity are also critical factors to consider before any further experimental exploration of samples should be pursued.
Day 1 was rounded off with a boat trip along the Thames. It was a clear and beautiful September evening and everyone had a fantastic view of the famous sites of London, including the Houses of Parliament and Tower Bridge. This was a great finish to a day of fantastic and stimulating workshops, and was the perfect opportunity to network with all the delegates.
Ronan O’Brien, MicroCal’s Head of Business Development, kicked off another glorious day in London, with lots of excitement about the fantastic scientific agenda ahead of us. The first session of the morning was focused on ITC, with Geoff Holdgate of AstraZeneca sharing his use of ITC in the drug discovery process, highlighting the parameters his team uses in their workflow. ITC has helped them obtain structural information, along with determining the binding mechanism for their compounds. The next presentation by James Murray of Vernalis showed how ITC was being used in his team’s screening rationale, to screen fragments in their crude reactions as well as in cleaner experiments. ITC has become an important part of their workflow for confirming hits, but also enabling them to obtain a complete thermodynamic and kinetic profile from crude mixtures. Eric Ennifar of CNRS Strasbourg University explained how he is using the Malvern Panalytical PEAQ-ITC to study ribosome binding to antibodies and viral RNAs, obtaining kinetic data from these experiments. Five posters were selected by the Scientific Committee of the meeting for flash poster presentations by Inna Rozman Grinberg from Stockholm University, Sweden, Jascindra Ravi from National Physical Laboratory in Teddington, UK, Sergei Kuprin from Swedish Orphan Biovitrum AB, Stockholm, Sweden, Marie Markones from the University of Freiburg, Germany. Robert Risti from Tallinn University of Technology, Estonia and Katharina Weickhmann from Goethe Universität Frankfurt, Germany. A panel discussion, chaired by Mikael Akke of Lund University, closed off the first morning session. There was great interaction from the audience and lots of discussion amongst the delegates and panelists, resulting in a great buzz in the auditorium.
The second session of the morning, led by Michaela Wimmerova of CEITEC, was focused on DSC, and we were treated to some fantastic talks from a range of customers. Thomas Meier of Roche highlighted the value of DSC within his workflow for lot-to-lot comparison, and also for process and buffer optimization. Marina Kasimova of Symphogen provided a very enthusiastic account of how she used DSC in stabilization studies of the Fc domain of IgG1. The final talks included the use of DSC in the PPC mode for looking at protein systems, which was presented by Mariano Dellarole of the Institut Pasteur. Peng Ke of MedImmune rounded off the presentations, highlighting the use of DSC as part of their antibody development as part of an orthogonal approach. There was great engagement between the panelists, and Chris Johnson of the Medical Research Council was able to add extensive knowledge to this subject matter. Lots of great questions were put forward by the delegates, finishing off a fantastic morning session.
During lunch, delegates had the opportunity to network and absorb the great posters that were on display. These came from many countries across Europe and had lots of exciting content covering all aspects of calorimetry. Discussions were in full flow and it was great to see young scientists engaging with very experienced scientists from across industry and academia sharing their knowledge and expertise. Delegates had the opportunity to see the new Zetasizer Ultra in action – this is a valuable tool for providing details of protein integrity prior to techniques such as ITC and DSC.
Bertrand Raynal from the Institut Pasteur kicked off the first of the afternoon sessions (focused on data quality) with a very energetic presentation about the importance of determining standardized protein sample quality prior to any further biophysical characterization. Applying a QC protocol provides better data quality, and will save time and money by enabling the upfront determination of sample aggregation or contamination. Steffen Glöckner from Philipps-University Marburg presented “Novel data on how kinITC and high-resolution macromolecular crystallography can be combined to establish structure-thermodynamic and -kinetic relationships to facilitate drug development”. Anders Dybdal Nielsen of Novo Nordisk followed with an interesting account of how his team is using PEAQ-ITC to measure the binding of the growth hormone somapacitan and human albumin.
We were then treated to a third panel discussion, led by Stephan Uebel, about the importance of having good quality samples and good procedures to ensure the best outcomes prior to techniques like ITC or SPR.
The final session of the meeting was focused on developments in the field of ITC, and was chaired by Eric Ennifar. Adrian Velazquez- Campoy shared work that was conducted with ARBRE-MOBIEU on an inter-laboratory study carried out using different models of MicroCal ITC systems and 4 different versions of software, with a view to assessing precision and accuracy and comparing data handling. This study has helped with the recommendation of test reactions for use in ITC data standardization. Ksenia Maximova of Warsaw University spoke enthusiastically of her use of ITC as a reliable label-free technology to measure the kinetic parameters of enzymatic reactions. Eva Muñoz rounded off the formal presentations, with a presentation of a case study, in which she stated how ITC was considered the gold standard for thermodynamic characterization but could also be used for kinetic characterization with the Affimeter software.
To finish the meeting, the fourth and final panel discussion provided a focus on the analysis of complex ITC data, which sparked further discussion and interaction between the delegates and experts on the panel. All four panel discussions provided a great forum for questions, interesting and thought-provoking discussion and considerations for the calorimetry community, with particular emphasis on how to obtain good quality meaningful data from the MicroCal instruments. Natalia Markova closed the meeting by summarizing the key take-home messages.
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I would like to thank all the speakers, panelists, poster presenters, scientific committee members, and of course all the delegates who attended this meeting, making it an exciting, dynamic and informative event. Roll on the next meeting!