| 00:00:00 | Welcome |
| 00:00:13 | Introduction |
| 00:00:58 | Revealing the secrets of nanodisc architecture using multi-detection GPC/SEC |
| 00:01:11 | Outline |
| 00:02:19 | Motivation |
| 00:03:36 | Motivation |
| 00:03:56 | Motivation |
| 00:05:43 | Motivation |
| 00:07:25 | Theory: Omnisec |
| 00:07:34 | OmniSEC |
| 00:08:28 | Untitled |
| 00:09:14 | Concentration detectors(RI and UV absorption) |
| 00:10:47 | Molar mass detector(static light scattering) |
| 00:12:57 | Molecular size detector(viscometry) |
| 00:14:21 | Molecular size detector(dynamic light scattering) |
| 00:15:11 | Results: BSA and SMALPs |
| 00:15:24 | Experimental conditions |
| 00:16:50 | BSA chromatogram |
| 00:17:41 | BSA analysis |
| 00:18:49 | BSA analysis |
| 00:19:27 | SMALP analysis(method) |
| 00:20:23 | SMALP chromatogram |
| 00:22:36 | SMALP analysis (SMA distribution) |
| 00:23:41 | SMALP analysis (SMA distribution) |
| 00:24:47 | SMALP analysis (SMA distribution) |
| 00:25:47 | SMALP analysis (SMALP and SMA molar mass) |
| 00:26:25 | SMALP analysis (SMALP size and composition) |
| 00:28:10 | SMALP analysis (SMALP size and shape) |
| 00:31:00 | Summary and conclusions |
| 00:32:05 | Acknowledgements |
| 00:32:35 | Q&A |
| 00:39:12 | Contact Information |
Membrane protein research is dependent on the availability of membrane mimics that retain the structure and function of the target protein within a lipid-bilayer environment while rendering it amenable to in vitro investigations.
Styrene/maleic acid (SMA) copolymers extract phospholipids and proteins from artificial and natural membranes to form polymer-bounded nanodiscs (SMALPs) which retain the lipid-bilayer architecture of the host membrane. Therefore, SMALPs are gaining increasing interest among membrane-protein researchers as an attractive alternative to conventional membrane mimics such as micelles and liposomes. However, the morphology of SMALPs, and, in particular, the extent to which they provide a “protein-friendly” lipid-bilayer environment, remains poorly characterized.
In this webinar, we will demonstrate how multi-detector GPC/SEC provides compositional and morphological information on SMALPs, contributing to a better, quantitative understanding of these new membrane-mimetic nanoparticles.
演講者
Johannes Klingler studied Biophysics at the University of Kaiserslautern and received his Diploma in 2013 as a member of Prof. Dr. Sandro Keller’s lab group. Since then, he has worked within the same group on his PhD project, which is focused on the structural and thermodynamic characterization of polymer/lipid nanoparticles by spectroscopic, calorimetric, chromatographic, and light scattering techniques.