Accurate characterization of biomacromolecules is essential for successful programs of research and development in the life sciences and biopharmaceuticals. The basic biophysical properties of these molecules include molecular weight, size, conformation, degree of conjugation, aggregation and complex-forming interactions.

Size-exclusion chromatography is commonly used to separate and analyze proteins and other biomacromolecules. However, in order to reliably determine their basic biophysical properties in solution, an absolute, independent means of characterization must be added downstream of the separation step.

Multi-angle light scattering and dynamic light scattering instruments, combined with UV and RI detectors, fulfill that need, making them essential in every lab that produces, uses or characterizes proteins, peptides, nucleic acids, polysaccharides or bionanoparticles constructed of these building blocks. This article explores the technology, capabilities and applications of light scattering paired with size-exclusion chromatography for biophysical characterization.

DAWN®

DAWN®

The world’s most advanced light scattering instrument for absolute characterization of proteins, conjugates, macromolecules, and nanoparticles.

The DAWN and its companion Optilab dRI detector are the established benchmarks for MALS analysis, cited in thousands of peer-reviewed publications. Multi-angle light scattering detection is indispensable for use with GPC and HPLC-SEC in order to obtain reliable molecular mass distributions and information on molecular conformation, branching ratio, fragments and aggregates.

Optilab

Optilab

Universal Detection for Chromatography and FFF Separations

Using a combination of cutting-edge semiconductor photodiode technology and proprietary computer algorithms, the Optilab achieves an unprecedented combination of sensitivity and range. These features mean that it addresses both standard chromatographic applications and some challenges unique to light scattering measurements such as high concentrations, determination of sample refractive increments (dn/dc) and solvent refractive index.

ViscoStar

ViscoStar

ViscoStar: The next generation online differential viscometer.

The ViscoStar® incorporates multiple novel technologies to provide the highest sensitivity, stability and solvent compatibility of any available viscometer for GPC. Its ease-of-use and serviceability make it the perfect companion for Wyatt’s DAWN® light scattering and Optilab® refractive index detectors.