Wyatt Technology

Analyzing Ebola virus glycoprotein and its interactions with therapeutic antibodies using CG-MALS

 

Speakers:
Dr. Kathryn Hastie, The Scripps Research Institute
Dr. Daniel Some, Principal Scientist

Presented Live: March 15, 2017

The Ebolavirus (EBOV) glycoprotein (GP) is essential for the attachment and fusion of virus and host cell membranes. It is also the primary target of neutralizing and therapeutic antibody responses. EBOV GP is 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 EBOV GP and antibodies that are neutralizing or protective. Using this technique we were able 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 (Pallesen, et al. Nature Microbiology, 2016).

This webcast will explain how we characterize the glycan content and oligomerization state of EBOV GP 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.

 

 

   

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Wyatt Technology is the recognized leader in light scattering instrumentation and software for determining the absolute molar mass, size, charge and interactions of macromolecules and nanoparticles in solution.

Wyatt's line of multi-angle static light scattering products couple to size exclusion chromatography (SEC-MALS), field-flow fractionation (FFF-MALS), and stop-flow composition-gradient systems (CG-MALS). Our dynamic light scattering (DLS) products operate in traditional cuvette as well as on-line and automated, high-throughput modes. We also offer unique instruments for electrophoretic light scattering (MP-PALS), differential refractometry, and differential viscosity.



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