Seznámení s přístroji Wyatt Technology

Significance of the Topic

Multi-angle light scattering (MALS), asymmetric flow field-flow fractionation (AF4) and dynamic light scattering (DLS) have become essential analytical techniques for characterizing macromolecules and nanoparticles in solution. They provide absolute molar mass, size, conformation and interaction data critical for protein research, biopharmaceutical development, drug delivery systems, polymer science and industrial quality control.

Objectives and Study Overview

This presentation introduces Wyatt Technology’s suite of light scattering and separation instruments now integrated into Waters Corporation. It aims to outline instrument capabilities, methodologies, and application examples that demonstrate how these technologies enhance the analysis of molecular weight distributions, structural information and solution behavior.

Used Instrumentation

• DAWN and miniDAWN Multi-Angle Light Scattering (MALS) detectors for absolute molar mass and radius of gyration
• Optilab Refractive Index Detector for concentration measurements
• ViscoStar Online Viscometer for intrinsic viscosity and hydrodynamic radius determination
• Eclipse AF4 module with dilution control for asymmetric flow field-flow fractionation, minimizing shear degradation
• NanoStar, PlateReader and ZetaStar Dynamic Light Scattering (DLS) instruments for hydrodynamic radius, diffusion coefficient, zeta potential, concentration and turbidity analysis

Main Results and Discussion

The coupling of SEC or AF4 with MALS yields absolute molecular weight distributions and size distributions without relying on column calibration. SEC-MALS can determine molar mass, radius of gyration and branching via the slope of log–log plots of radius versus molar mass. Combined viscometry adds conformational insight through intrinsic viscosity measurements. AF4-MALS overcomes polymer anchoring and shear degradation in chromatography, extending analysis to higher mass ranges. DLS provides rapid determination of hydrodynamic radii and diffusion coefficients, complementing MALS data. ZetaStar further integrates charge, concentration and stability assessments from microvolume samples.

Benefits and Practical Applications

• Absolute measurement of molar mass distributions, size, conformation and branching
• Reduction of analytical artifacts such as shear degradation and column interactions via AF4
• Simultaneous multi-parameter characterization in a single run (mass, size, viscosity, charge)
• High sensitivity and low sample consumption for biopharma, protein formulation and polymer QA/QC
• Versatile online coupling options enabling real-time analysis

Future Trends and Potential Applications

Advances in detector sensitivity, microfluidic integration and machine learning-based data analysis are expected to further enhance resolution and throughput. Emerging applications include characterization of complex coacervates, nanoparticles, drug delivery vehicles and structural studies of intrinsically disordered proteins. Integration with hyphenated techniques such as mass spectrometry and microfluidics will expand the analytical capabilities for next-generation materials and therapeutics.

Conclusion

Wyatt Technology’s light scattering and separation platforms provide a comprehensive toolbox for absolute macromolecular characterization. Their integration into Waters accelerates access to high-resolution molecular data crucial for research and quality control in diverse fields ranging from polymer science to biopharmaceuticals.

References

• Podzimek S, Vlcek T, Johann C. J Appl Polym Sci. 2001;81:1588.