Universal building blocks for antigen presentation to uncover the flavivirus-dir...
Universal building blocks for antigen presentation to uncover the flavivirus-directed antibody response
Flaviviruses are a tremendous burden on global public health. The genus consists of yellow fever virus, dengue virus, and zika virus amongst others. These viruses threaten more than half the global human population and are spreadi...
Flaviviruses are a tremendous burden on global public health. The genus consists of yellow fever virus, dengue virus, and zika virus amongst others. These viruses threaten more than half the global human population and are spreading into new territories. The recent zika virus pandemic stresses the need for a suitable protein engineering toolkit to enable swift development of emerging flavivirus antigens for structural studies, serology and vaccine development. Moreover, the structural determinants of a neutralizing polyclonal antibody response are currently not well understood, in large part for a lack of suitable methods to address the complex interactions of multiple antibodies directed against a heterogeneous, glycosylated viral antigen.
With FLAVIR I will use my unique expertise in both mass spectrometry and electron microscopy to develop a comprehensive structural proteomics toolkit for in-depth profiling of the neutralizing antibody response against flaviviruses. Using a combination of novel mass spectrometry-based antibody sequencing techniques, glycoproteomics profiling and electron microscopy, we will uncover how the polyclonal antibody response against flaviviruses is directed by key structural determinants of the Envelope glycoprotein that is displayed on the surface of infectious virions.
We will develop new strategies to produce prefusion stabilized E-dimers and ultimately reconstruction of full icosahedral flavivirus-like particles from soluble components. The outlined strategies will be tested and streamlined for broad applicability across the flaviviruses so that it can be swiftly adapted for emerging species and strains. We will characterize our designs with state-of-the art mass spectrometry methods (native MS, mass photometry, HDX-MS) and electron microscopy.
We will use these optimized antigens to uncover the role of antigen glycosylation in antibody binding, profile the serum antibody repertoire directed against the E-antigen and map ouver más
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