| 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.