記録された日時: July 17 2018

Duration: 45 minutes 13 seconds

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.
Table of contents
1. Welcome
00:19
2. Introduction
01:14
3. Title
00:56
4. Objective and Outline
01:29
5. Rayleigh light scattering equation gives direct measure of molar mass
06:01
6. 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)
01:57
7. Precision and limits of detection of SEC-SLS1
06:18
8. For a simple polymer (approximately uniform atomic composition) 2 detectors are sufficient for characterization
01:06
9. Size estimates for proteins
03:39
10. Debye Plot gives radius of gyration
01:20
11. 2-component system non-ideality
03:11
12. PEGylated peptide case study 1 (5kDa PEG with ~ 5.8kDa peptide): Degree of PEGylation
05:11
13. 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
00:50
14. PEGylated peptide case study 2 (40kDa PEG with 2 × 2kDa peptides): Mw and Size Determination with Malvern OMNISEC system
04:30
15. OMNISEC analysis reveals that addition of peptides to both ends of PEG results in significant compaction of size
03:02
16. Acknowledgements
00:16
17. Thank you for your attention
03:42
18. Contact Information
00:12