Dynamic light scattering: a practical guide and applications in biomedical sciences

Abstract

Dynamic light scattering (DLS), also known as photon correlation spectroscopy (PCS), is a very powerful tool for studying the diffusion behaviour of macromolecules in solution. The diffusion coefficient, and hence the hydrodynamic radii calculated from it, depends on the size and shape of macromolecules. In this review, we provide evidence of the usefulness of DLS to study the homogeneity of proteins, nucleic acids, and complexes of protein-protein or protein-nucleic acid preparations, as well as to study protein-small molecule interactions. Further, we provide examples of DLS's application both as a complementary method to analytical ultracentrifugation studies and as a screening tool to validate solution scattering models using determined hydrodynamic radii.

Detection of Small‐Molecule Aggregation with High‐Throughput Microplate Biophysical Methods

“…DLS measures the fast (nanosecond) fluctuations of scattered light intensity by particles in solution as they diffuse due to Brownian motion, and determines the translational diffusion coefficient. The translational diffusion coefficient can be used to calculate the particle hydrodynamic radius (R h ) and size using equations derived from the first principles of classical physics (Stetefeld, McKenna, & Patel, 2016). Scratches on the glass bottom of microwell plates and dust particles cause large and artifactual light scattering effects.…”

Probing Polymer Chain Conformation and Fibril Formation of Peptide Conjugates

“…[5] A large variety of experimental and theoretical methods has been used in the past to investigate the shape and form of unfolded macromolecules, such as static-and dynamic light scattering [6] or viscosimetry, dating back to the fundamental investigations of Staudinger. [7] Modern methods to probe e. g. distance-relations between the ends of macromolecules including methods such as fluorescence resonance energy transfer (FRET), [8] triplet/triplet-energy transfer (TTET), [9] or ESR-spectroscopy [10] provide an average picture of chain extension. Especially the largely growing field of singlechain folded macromolecules, [5,11] where an individual chain of a synthetic macromolecule is folding into a singular nanoparticle, [12] is still elusively explored, because of a lack of specific methods to probe the folding of individual polymer based nanoparticles.…”

Establishing Trypanosoma cruzi farnesyl pyrophosphate synthase as a viable target for biosensor driven fragment‐based lead discovery

“…The quaternary structure and homogeneity of the purified protein was evaluated by dynamic light scattering (DLS). The technique estimates the molecular weight and size distribution of the protein via measurements of its diffusion in solution over time . A “distribution by volume analysis” of the protein (stock solution) indicated that there was essentially only a single species of protein present (Figure 2b).…”

“…The technique estimates the molecular weight and size distribution of the protein via measurements of its diffusion in solution over time. 18 A "distribution by volume analysis" of the protein (stock solution) indicated that there was essentially only a single species of protein present ( Figure 2b). The hydrodynamic diameter of the protein was estimated to be 7.14 nm, corresponding to the molecular weight of a dimer (84 KDa).…”