Applications of light scattering and intrinsic viscosity detectors with size exclusion chromatography (SEC) in characterizing biopharmaceutical and other polymer products

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00:00:00 Welcome
00:00:19 Introduction
00:01:33 Title
00:02:29 Objective and Outline
00:03:58 Rayleigh light scattering equation gives direct measure of molar mass
00:09:59 dn/dc of the protein will depend on the solvent it is dissolved in (dn/dc is one of the greatest sources of error in SLS measurements)
00:11:56 Precision and limits of detection of SEC-SLS1
00:18:14 For a simple polymer (approximately uniform atomic composition) 2 detectors are sufficient for characterization
00:19:20 Size estimates for proteins
00:22:59 Debye Plot gives radius of gyration
00:24:19 2-component system non-ideality
00:27:30 PEGylated peptide case study 1 (5kDa PEG with ~ 5.8kDa peptide): Degree of PEGylation
00:32:41 Using on-line SEC-SLS detection, with (dn/dc)PEG-protein = 0.147 ml/g from UV-dRI, enables accurate molecular weight determination of a PEGylated peptide
00:33:31 PEGylated peptide case study 2 (40kDa PEG with 2 × 2kDa peptides): Mw and Size Determination with Malvern OMNISEC system
00:38:01 OMNISEC analysis reveals that addition of peptides to both ends of PEG results in significant compaction of size
00:41:03 Acknowledgements
00:41:19 Thank you for your attention
00:45:01 Contact Information
This webinar will provide a quick introduction to the basics of UV, dRI, MALS, and intrinsic viscosity detection for size exclusion chromatography (SEC).  Applications covered will include the determination of molecular weight by UV or dRI (differential refractive index) and MALS (multi-angle light scattering),  measurement of the degree of PEGylation with UV, dRI and MALS, and elucidation of structural and hydrodynamic properties using IV (intrinsic viscosity) and MALS.