Analysis of Ebola Virus Glycoproteins in Complex with Antibodies using Light Scattering
Viral glycoproteins are essential for the attachment and fusion of virus and host cell membranes. It is also the primary target of neutralizing and therapeutic antibody responses. Many viral GPs are found as a trimeric complex on the viral surface. Whether multiple antibodies bind to these complexes and with what affinity is critical information when designing vaccines or creating a post-exposure antibody treatments.
Using composition-gradient multi-angle light scattering (CG-MALS), which quantifies the change in apparent weight-average molar mass (Mw) as a function of composition, we analyzed the reversible association between the viral glycoproteins of Ebola virus and Lassa virus and antibodies that are neutralizing or protective. This technique enabled us to determine the affinity and stoichiometry of various immune complexes as well as the relative quantity of each type of complex. The best-fit model for the CG-MALS data was supported by structural characterization using cryoEM.
This webcast will explain how we characterize the glycan content and oligomerization state of multimeric viral glycoproteins using SEC-MALS and demonstrate the detailed information we can gain about GP-antibody interactions using CG-MALS. The webinar will address the preparation of proteins for light-scattering studies as well as the set-up of the instruments and analysis of the data. This webinar will enable users of Wyatt light scattering instruments to set up and perform their own SEC-MALS and CG-MALS experiments and critically evaluate the resulting data.
Key Learning Objectives:
- Understanding the latest Ebola virus treatment and research.
- Absolute mass analysis of glycoproteins using SEC-MALS.
- Affinity and stoichiometry of antibody binding to multivalent glycoproteins.
Presenter: Kathryn Hastie (Instructor and Director of Antibody Discovery)
Dr. Hastie uses protein engineering and high-resolution structural analysis to design better therapeutics and vaccines against viruses that threaten global health. She is a leading expert on viral glycoproteins and their antigenic landscapes, marrying structural insights and vaccine design. Dr. Hastie further spearheads antibody discovery efforts at La Jolla Institute for Immunology to find antibodies against emerging viruses and other therapeutic targets.
