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Quickly Screen Peptides, Proteins and Nanoparticles for Size and Aggregation Using Advanced Methods

 

Presented Live: November 14, 2017

Dynamic Light Scattering (DLS) is the most widely employed technique for size and polydispersity measurement in protein, polymer, and nanoparticle research. Traditionally, DLS has been limited to measuring a single sample at one time in a cuvette, but the DLS Plate Reader from Wyatt breaks through this limitation by measuring entire well plates of samples at one time. This expanded measurement capacity combined with small volume requirement and flexible, custom experiment design enable analyses that were not previously possible.

 

  Webinar Q & A

 

 

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

Speakers:
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.

 

 

Small, but bright: µSEC-MALS adds light scattering to UHP-SEC for biophysical characterization of biotherapeutics

 

Speakers:
Dr. Stephan Koza, Principal Applications Chemist, Waters Corp.
Dr. Michelle Chen, Director of Analytical Services, Wyatt Technology Corp.
Mark Haverick, Senior Scientist, Merck & Co Inc.

UHP-SEC-MALS combines the high speed, high resolution, and low sample consumption of UHP-SEC separation with online multi-angle light scattering (MALS) analysis. MALS is the recognized, indispensable method for obtaining absolute molecular weight and size, independently of column retention time. In this webinar we present the key features of UHP-SEC as well as a MALS detector optimized to couple with it, creating a “µSEC-MALS” system for protein and synthetic polymer characterization.

Case studies will illustrate the applications of µSEC-MALS in the analysis and characterization of biopharmaceuticals. In particular we will discuss some of the challenges of bridging analytical-scale SEC-MALS to µSEC-MALS, as well as bridging between different UHP-SEC elution conditions.

 

 

   

For essential macromolecular and nanoparticle characterization—The Solution is Light™

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Wyatt Technology is the recognized leader in light scattering instrumentation and software for determining the absolute molar mass, size, charge and interactions of macromolecules and nanoparticles in solution.

Wyatt's line of multi-angle static light scattering products couple to size exclusion chromatography (SEC-MALS), field-flow fractionation (FFF-MALS), and stop-flow composition-gradient systems (CG-MALS). Our dynamic light scattering (DLS) products operate in traditional cuvette as well as on-line and automated, high-throughput modes. We also offer unique instruments for electrophoretic light scattering (MP-PALS), differential refractometry, and differential viscosity.



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