Protein structure is tightly linked to protein function. Detailed characterization of protein structure enables understanding and control of protein function and is therefore among the activities central to academic and industrial research and development.
The use of ever-more structurally complex molecules warrants a growing requirement for complementary and orthogonal analytics to ensure data quality and the reliability of research. The ‘first principle’ nature and high resolution of Differential Scanning Calorimetry (DSC) makes it a well-established technique for extended structural characterization and stability profiling of biomolecules and viruses in solution. Due to its direct readout, broad temperature range and sensitivity to thermally-induced unfolding, DSC is also used as the gold standard technique for validation of data from higher throughput thermal stability assays.
This webinar by David Staunton covers the basic principles of DSC and its application to the development of Neisseria meningitidis vaccine candidates.
Neisseria meningitidis is the leading cause of bacteraemia and sepsis in children and young adults due to the non-specific nature of its initial symptoms and the rapid progress of infection. Vaccination is the best approach to protect individuals and progress has been made in their development. However, the existing strategy cannot be used with serogroup B, which is the most common form of the disease in Europe and North America.
An alternative vaccine candidate is the factor H binding protein (fHbp), a 27 kDa lipoprotein that can be divided into three variant groups, V1, V2, and V3, and which consists of two beta barrel modules. Members of the variant groups share 85% amino acid identity, but only 60-70% similarity between groups and immunization with only one variant generates immunological cross-reactivity within but not between variant groups. Efforts have been made to design functionally inactive but immunogenic fHbp as vaccine candidates. X-ray structures of V1 and V3 were obtained, but only the carboxy beta barrel module was observed for V2. Differential Scanning Calorimetry (DSC) was used to determine the folding stability of the fHbp variants to explain this observation. The advantages of applying DSC over other protein stability analyses will be discussed.
- April 08 2020 - April 08 2020
10:30 - 11:30
(GMT-05:00) Eastern [US & Canada]
- Webinar - Live
Michael Caves - Pharmaceutical Segment Manager - Asia Pacific and Prof. David Staunton - Biophysics Suite, University of Oxford
Dr Michael Caves has spent 20 years working in various academic and commercial life science settings. He has published scientific papers on biotherapeutic characterisation and formulation, and has applied this knowledge during various roles within the pharmaceutical and diagnostics industries. Having helped build the formulation development service at SGS Life Sciences, Michael joined Malvern Panalytical in 2013 in order to support customers around the world. Michael is currently based in Chennai, India, from where he focuses on helping customers across Asia’s pharmaceutical industry.
David Staunton received his DPhil in virology from the University of Oxford in 1986, after which he held postdoctoral positions in Bath and Oxford, specializing in biophysical analyses on cytokines and their receptors, and fibronectin. He was appointed Facility Manager of the Biochemistry Department’s Molecular Biophysics Suite in 2008 and University Research Lecturer in 2010.
- Who should attend?
- Scientists and project leaders in academia and industry working with the characterization of proteins and the development of protein-based vaccines and therapeutics.
- What will you learn?
- Basic principles of DSC and its application to vaccine development
- Ways to apply DSC to the characterization of recombinant multi-domain proteins
- Benefits of DSC as compared to other thermal shift techniques