engleski

Advanced characterization of IgG formulations by multi-detection size exclusion chromatography

Size Exclusion Chromatography (SEC) coupled with multi-detector consisting of refractive index (RI), UV/Vis absorbance, light scattering (LS) and intrinsic viscosity (IV) detectors has already been recognized as a valuable analytical technique for the characterization of macromolecules as well as monitoring of the process control, manufacturing and formulation of biotechnology products. It provides deep insight into sample’s absolute molecular weight, molecular size (hydrodynamic size and radius of gyration), intrinsic viscosity, but also informations concerning structure, concentration, recovery, and potentially, absorption profile and compositional analysis. The aim of this work was to investigate the possibilities of the multi-detection size exclusion chromatography in advanced characterization of IgG-based antivenoms obtained by different purification principles. The absolute molecular weight (Mw) of the sample peaks, their hydrodynamic radius (Rh) and polydispersity (Mw/Mn) were determined. Trace amounts of aggregates and the differences in molecular nature of the polymeric structures (dimer, trimer, etc.) were detected. The obtained absolute Mw values were in line with the expected ones for dimeric and monomeric IgG species. Polydispersity value of ~1 confirmed molecular homogeneity and symmetrical size distribution for monomers and dimers. On the contrary, the peaks that correspond to soluble aggregates showed higher polydispersity values, indicating their heterogenous composition. This methodology proved as promising and powerful technology for the quality control of therapeutic IgGs during the process development.

Size Exclusion Chromatography ; advanced characterization ; multi-detection ; therapeutic IgG ; quality control

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