New Customer Application Note Available from Wyatt Technology

Wyatt Technology Corporation, the world leader in absolute macromolecular characterization instrumentation and software, publishes new application note highlighting a study made by Florida State University. Authored by Tsz Kin Martin Tsui, Claudius Mundoma and Hong Li from Florida State University, the paper showcases how the Wyatt DynaPro dynamic light scattering (DLS) system can be used to characterize homogeneity and stability of a heterodimeric protein complex (HPC) under various buffer conditions. The results demonstrate that the pH of the buffer solutions influences the sample homogeneity and stability.

A heterodimeric protein complex (HPC) from Pyrococcus furiosus is identified for its functional role in binding RNA. HPC has a total molecular weight of 65 kDa and has a tendency to form fibril materials in vitro as observed by electron microscopy. DLS data were acquired at 20°C using the Wyatt DynaPro NanoStar Dynamic Light Scattering (DLS) system. A total of 10 measurements were performed for each sample, and the stability and homogeneity determined from the polydispersity and autocorrelation functions using DLS provided information on the biophysical property of the HPC.

Hong Li, Department of Chemistry and Biochemistry and the Institute of Molecular Biophysics, Florida State University, said “Biological macromolecules are the focus of the Institute’s research – their form, function, interactions and mechanism of action. Challenging frontier areas include characterizations of the interactions between proteins and membranes, sugars or nucleic acids. The DynaPro NanoStar is ideally suited for studies of these complexes in addition to nanoparticles, vesicles, viruses and colloids. The results from the study demonstrate that the pH of the buffer solutions influences the sample homogeneity and stability, highlighting the conditions where polymerization of HPC is thermodynamically more favorable.

The Institute of Molecular Biophysics has a long history as a unique interdisciplinary research unit dedicated to the application of physical and chemical principles to the investigation of biological phenomena. Dr. Claudius Mundoma is the Director of the Physical Biochemistry facility which houses a broad array of biophysical technology used for macromolecular characterization of structure of macromolecules. He comments, “a technique such as Dynamic Light scattering is very efficient, requiring less time and sample than other techniques to determine particle size distribution and aggregation states of molecules.â€