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.
The need for biophysical characterization
Reliable analysis of the molecular weight (MW) of proteins in solution is essential for biomolecular research1–4. MW analysis informs the scientist if the correct protein has been obtained and if it is suitable for use in further experimentation5,6. As described on the web sites of protein networks P4EU7 and ARBRE-Mobieu8, protein quality control must characterize not only the purity of the final product, but also its oligomeric state, homogeneity, identity, conformation, structure, post-translation modifications and other properties.
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