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Absolute Characterization of Glycoproteins and their Interactions with Proteins and Antibodies by Light Scattering

 

Presented Live: September 6, 2017

Speakers: Dan Some, Ph.D.

Glycoproteins (GPs) are primary targets of neutralizing and therapeutic antibodies for a large variety of indications including epilepsy and many types of viruses. Many non-IgG therapeutic proteins are themselves GPs. Hence, the characterization of GPs and potential degradants, as well as their interactions with therapeutic or target proteins, is essential for understanding GP targets and developing effective therapeutics. This webinar presents two first-principles techniques for carrying out these analyses in solution: SEC-MALS-UV-RI and CG-MALS.

Unlike common unmodified proteins that are globular in conformation and highly hydrophilic in surface chemistry, GPs cannot be characterized accurately by standard analytical size-exclusion chromatography (SEC). Coupling SEC to multi-angle light scattering (MALS) and differential refractive index (dRI) detection results in a powerful SEC-MALS-UV-RI triple-detection system that can determine the molar mass of the protein and total glycan mass in order to characterize degradants and complexes. Composition-gradient multi-angle light scattering (CG-MALS) utilizes the same detectors with a stop-flow system to quantify the absolute stoichiometry and binding affinity of reversible association between GPs and antibodies or solubilized target proteins.

 

 

 

   

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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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