Applications of Analytical Light Scattering in a Biophysics Core Facility
Presented Live: December 5, 2017
Speakers: Ewa Folta-Stogniew, Ph.D.
Static light scattering (SLS) is an analytical technique for determination of molar masses (molecular weights) and radii of gyration of macromolecules in solution. Measurement of molar mass from an SLS experiment determines the oligomeric state and thus provides association stoichiometry for biological macromolecules such as native and modified proteins and their complexes.
This webcast presents several case studies of the application of size exclusion chromatography (SEC) coupled with multi-angle (static) light scattering (MALS) in analysis of protein structure-function relationship, carried out at the Biophysics Resource of Keck Biotechnology Research Laboratory at Yale University School of Medicine. The examples include studies of heparin-induced dimerization of anaplastic lymphoma kinase (ALK), determination of stoichiometry of nucleic-acid repressor complex involved in regulation of transcription of the c-myc oncogene, deciphering the pathway of oligomerization and higher-order assembly of PopZ polar scaffold protein that supports asymmetric cell division, and re-examination of the role of amino-terminus in dimerization of SecA protein.
Innovations and Applications of UHP-SEC and Multi-Angle Light Scattering (MALS) for the Analysis of Biotherapeutic Proteins
Presented Live: October 25, 2016
Dr. Stephan Koza, Principal Applications Chemist, Waters Corp.
Dr. John Champagne, Senior Applications Scientist, Wyatt Technology Corp.
Ultra-high performance size exclusion chromatography (UHP-SEC) is gaining in popularity owing to the high speed and resolution of separation compared to standard HP-SEC. Online multi-angle light scattering (MALS) is the recognized method for obtaining absolute molecular weight and size, independently of column retention time, making it indispensable for accurate and reliable characterization by SEC. However, light scattering detectors designed for HPLC-SEC are in general not suitable for UHP-SEC.
In this webinar we discuss the practical application of UHP-SEC with regards to method development and successful implementation. This will be followed by a discussion on limitations of standard MALS detectors and present the capabilities of a MALS detector designed explicitly to meet the challenges of UHP-SEC. The utility of UHP-SEC combined with MALS–dubbed ‘µSEC-MALS’–as well as online dynamic light scattering will be demonstrated in the context of protein characterization.